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写真a

ヒロセ トモノリ
廣瀬 智威
Tomonori Hirose
所属
医学研究科 医科学専攻 分子生物学 講師
医学部 医学科
職名
講師
プロフィール

大学院時代から一貫して、細胞極性制御因子PAR3やaPKCλ/ζの哺乳類における機能解析を進め、現在は神経幹細胞の増殖・分化バランスを調整する分子機構解明を進めています。

・PAR3-aPKC複合体が細胞極性制御において重要な機能を持つことを解明。
(培養上皮細胞を用いたPAR3の機能解析)
・PAR3による細胞極性制御を必要とする形態形成過程の具体例を確認。
(マウス心臓発生過程でPAR3による細胞極性制御の必要性を提示)
・腎疾患の病態に細胞極性異常が関わる可能性を提唱。
(腎糸球体ポドサイト特異的aPKCλノックアウトにより、巣状糸球体硬化症モデルマウスの樹立に成功)
・PAR3による神経幹細胞の増殖・分化バランスを調整する新しい分子機構を解明。
(終脳特異的PAR3ノックアウトマウスにより、神経分化開始時に神経幹細胞の増殖を制限するための分子機構を提示)

・共同研究:表皮幹細胞の維持・分化・癌化における細胞極性制御因子の機能解明。
(表皮特異的aPKCλノックアウトにより、aPKCλによる表皮幹細胞の休止期制御機構を提示表非特異的PAR3ノックアウトと化学発癌モデルを組み合わせ、組織型によるPAR3依存性の違いを提示。)
・共同研究:血管形成・動脈疾患病態における細胞極性制御因子の機能解明。
(内皮細胞特異的PAR3ノックアウトや動脈硬化症モデルとの組み合わせにより、PAR3によるVEGF受容体の動態制御機構や動脈硬化・炎症抑制機構を提示)

外部リンク

学位

  • 医学博士 ( 横浜市立大学 )

研究キーワード

  • 細胞極性

  • 上皮細胞

  • 神経幹細胞

  • ヘッジホッグシグナリング

  • 一次繊毛

  • 糸球体疾患

  • 包括脳ネットワーク

  • PAR6

  • シグナル伝達

  • ASIP

  • aPKC

  • ASPP2

  • PAR3

  • ノックアウトマウス

  • 疾患モデルマウス

  • 心外膜前駆細胞

  • ポドサイト

  • 腎臓病

  • 細胞分化

  • 非対称分裂

研究分野

  • ライフサイエンス / 腎臓内科学  / 糸球体疾患

  • ライフサイエンス / 解剖学  / 発生学

  • ライフサイエンス / 病態医化学

  • ライフサイエンス / 発生生物学  / 神経発生

  • ライフサイエンス / 神経科学一般  / 神経幹細胞

  • ライフサイエンス / 分子生物学  / シグナル伝達

▼全件表示

所属学協会

  • 日本生化学会

    2015年7月 - 現在

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  • 日本分子生物学会

    1997年4月 - 現在

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委員歴

  • 横浜市立大学医学部医学科   医学教育分野別認証評価準備委員会  

    2015年4月 - 現在   

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    団体区分:その他

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論文

  • PAR3 restricts the expansion of neural precursor cells by regulating hedgehog signaling. 国際誌

    Tomonori Hirose, Sugitani Y, Hidetake Kurihara, Hiromi Kazama, Kusaka C, Noda T, Hidehisa Takahashi, Shigeo Ohno

    Development (Cambridge, England)   149 ( 21 )   2022年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:The Company of Biologists  

    ABSTRACT

    During brain development, neural precursor cells (NPCs) expand initially, and then switch to generating stage-specific neurons while maintaining self-renewal ability. Because the NPC pool at the onset of neurogenesis crucially affects the final number of each type of neuron, tight regulation is necessary for the transitional timing from the expansion to the neurogenic phase in these cells. However, the molecular mechanisms underlying this transition are poorly understood. Here, we report that the telencephalon-specific loss of PAR3 before the start of neurogenesis leads to increased NPC proliferation at the expense of neurogenesis, resulting in disorganized tissue architecture. These NPCs demonstrate hyperactivation of hedgehog signaling in a smoothened-dependent manner, as well as defects in primary cilia. Furthermore, loss of PAR3 enhanced ligand-independent ciliary accumulation of smoothened and an inhibitor of smoothened ameliorated the hyperproliferation of NPCs in the telencephalon. Thus, these findings support the idea that PAR3 has a crucial role in the transition of NPCs from the expansion phase to the neurogenic phase by restricting hedgehog signaling through the establishment of ciliary integrity.

    DOI: 10.1242/dev.199931

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    その他リンク: https://journals.biologists.com/dev/article-pdf/doi/10.1242/dev.199931/2232602/dev199931.pdf

  • The PAR3-aPKC-PAR6 complex 招待

    Shigeo Ohno, Spyros Goulas, Tomonori Hirose

    Cell Polarity 1: Biological Role and Basic Mechanisms   3 - 23   2015年1月

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    記述言語:英語   掲載種別:論文集(書籍)内論文   出版者・発行元:Springer International Publishing  

    The PAR3-aPKC-PAR6 complex (the aPKC-PAR complex) is among the most well-studied “polarity proteins” that play fundamental roles in cell polarity in a variety of biological contexts. It is one of the core signaling cassettes containing a protein kinase and scaffold proteins that is conserved in multicellular organisms. One of the most important features of “polarity proteins” is that many of them localize to specific sites on the cytoplasmic side of the cell periphery and position other polarity proteins through antagonistic interactions and positive feedback mechanisms. Asymmetric distribution of polarity proteins in the cell periphery thus provides cellular landmarks required for the generation and maintenance of cell polarity. Here, we describe what we know about the mechanisms of how the aPKC-PAR complex is specifically positioned and activated and regulates overall cell polarity of epithelial cells with special attention to its molecular nature.

    DOI: 10.1007/978-3-319-14463-4_1

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  • An Essential Role of the Universal Polarity Protein, aPKC lambda, on the Maintenance of Podocyte Slit Diaphragms 査読

    Tomonori Hirose, Daisuke Satoh, Hidetake Kurihara, Chiho Kusaka, Hiroko Hirose, Kazunori Akimoto, Taiji Matsusaka, Iekuni Ichikawa, Tetsuo Noda, Shigeo Ohno

    PLOS ONE   4 ( 1 )   e4194   2009年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    Glomerular visceral epithelial cells (podocytes) contain interdigitated processes that form specialized intercellular junctions, termed slit diaphragms, which provide a selective filtration barrier in the renal glomerulus. Analyses of disease-causing mutations in familial nephrotic syndromes and targeted mutagenesis in mice have revealed critical roles of several proteins in the assembly of slit diaphragms. The nephrin-podocin complex is the main constituent of slit diaphragms. However, the molecular mechanisms regulating these proteins to maintain the slit diaphragms are still largely unknown. Here, we demonstrate that the PAR3-atypical protein kinase C (aPKC) -PAR6 beta cell polarity proteins co-localize to the slit diaphragms with nephrin. Furthermore, selective depletion of aPKC lambda in mouse podocytes results in the disassembly of slit diaphragms, a disturbance in apico-basal cell polarity, and focal segmental glomerulosclerosis (FSGS). The aPKC-PAR3 complex associates with the nephrin-podocin complex in podocytes through direct interaction between PAR3 and nephrin, and the kinase activity of aPKC is required for the appropriate distribution of nephrin and podocin in podocytes. These observations not only establish a critical function of the polarity proteins in the maintenance of slit diaphragms, but also imply their potential involvement in renal failure in FSGS.

    DOI: 10.1371/journal.pone.0004194

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  • PAR3 is essential for cyst-mediated epicardial development by establishing apical cortical domains 査読

    T Hirose, M Karasawa, Y Sugitani, M Fujisawa, K Akimoto, S Ohnos, T Noda

    DEVELOPMENT   133 ( 7 )   1389 - 1398   2006年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Epithelial cysts are one of the fundamental architectures for mammalian organogenesis. Although in vitro studies using cultured epithelial cells have revealed proteins required for cyst formation, the mechanisms that orchestrate the functions of these proteins in vivo remain to be clarified. We show that the targeted disruption of the mouse Par3 gene results in midgestational embryonic lethality with defective epicardial development. The epicardium is mainly derived from epicardial cysts and essential for cardiomyocyte proliferation during cardiac morphogenesis. PAR3-deficient epicardial progenitor (EPP) cells do not form cell cysts and show defects in the establishment of apical cortical domains, but not in basolateral domains. In PAR3-deficient EPP cells, the localizations of aPKC, PAR6 beta and ezrin to the apical cortical domains are disturbed. By contrast, ZO1 and alpha 4/beta 1 integrins normally localize to cell-cell junctions and basal domains, respectively. Our observations indicate that EPP cell cyst formation requires PAR3 to interpret the polarity cues from cell-cell and cell-extracellular matrix interactions so that each EPP cell establishes apical cortical domains. These results also provide a clear example of the proper organization of epithelial tissues through the regulation of individual cell polarity.

    DOI: 10.1242/dev.02294

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  • Involvement of ASIP/PAR-3 in the promotion of epithelial tight junction formation 査読

    T Hirose, Y Izumi, Y Nagashima, Y Tamai-Nagai, H Kurihara, T Sakai, Y Suzuki, T Yamanaka, A Suzuki, K Mizuno, S Ohno

    JOURNAL OF CELL SCIENCE   115 ( 12 )   2485 - 2495   2002年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    The mammalian protein ASIP/PAR-3 interacts with atypical protein kinase C isotypes (aPKC) and shows overall sequence similarity to the invertebrate proteins C. elegans PAR-3 and Drosophila Bazooka, which are crucial for the establishment of polarity in various cells. The physical interaction between ASIP/PAR-3 and aPKC is also conserved in C. elegans PAR-3 and PKC-3 and in Drosophila Bazooka and DaPKC. In mammals, ASIP/PAR-3 colocalizes with aPKC and concentrates at the tight junctions of epithelial cells, but the biological meaning of ASIP/PAR-3 in tight junctions remains to be clarified. In the present study, we show that ASIP/PAR-3 staining distributes to the subapical domain of epithelial cell-cell junctions, including epithelial cells with less-developed tight junctions, in clear contrast with ZO-1, another tight-junction-associated protein, the staining of which is stronger in cells with well-developed tight junctions. Consistently, immunogold electron microscopy revealed that ASIP/PAR-3 concentrates at the apical edge of tight junctions, whereas ZO-1 distributes alongside tight junctions. To clarify the meaning of this characteristic localization of ASIP, we analyzed the effects of overexpressed ASIP/PAR-3 on tight junction formation in cultured epithelial MDCK cells. The induced overexpression of ASIP/PAR-3, but not its deletion mutant lacking the aPKC-binding sequence, promotes cell-cell contact-induced tight junction formation in MDCK cells when evaluated on the basis of transepithelial electrical resistance and occludin insolubilization. The significance of the aPKC-binding sequence in tight junction formation is also supported by the finding that the conserved PKC-phosphorylation site within this sequence, ASIP-Ser827, is phosphorylated at the most apical tip of cell-cell contacts during the initial phase of tight junction formation in MDCK cells. Together, our present data suggest that ASIP/PAR-3 regulates epithelial tight junction formation positively through interaction with aPKC.

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  • Biallelic TEDC1 variants cause a new syndrome with severe growth impairment and endocrine complications. 国際誌

    Noriko Miyake, Kentaro Shiga, Yuya Hasegawa, Chisato Iwabuchi, Kohei Shiroshita, Hiroshi Kobayashi, Keiyo Takubo, Fabien Velilla, Akiteru Maeno, Toshihiro Kawasaki, Yukiko Imai, Noriyoshi Sakai, Tomonori Hirose, Atsushi Fujita, Hidehisa Takahashi, Nobuhiko Okamoto, Mikako Enokizono, Shiho Iwasaki, Shuichi Ito, Naomichi Matsumoto

    European journal of human genetics : EJHG   2025年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We encountered two affected male patients born to non-consanguineous parents, who presented with prenatal-onset severe growth impairment, primary microcephaly, developmental delay, adrenal insufficiency, congenital glaucoma, delayed bone aging, craniosynostosis, congenital tracheal stenosis, and primary hypogonadism. By exome sequencing, we identified compound heterozygous TEDC1 variants (NM_001134877.1 c.[104-5C>G];[787delG] p.[?];[(Ala263LeufsTer29)] in both affected siblings. We confirmed that the splice site variant, c.104-5C>G, leads to no TEDC1 protein production via nonsense-mediated mRNA decay. The frameshift variant located in the last coding exon, c.787delG, produces a C-terminally truncated protein, which impairs the binding with TEDC2. Thus, both variants are thought to be loss-of-function. TEDC1 and TEDC2 are both required for centriole stability and cell proliferation. Our in vitro experiments using patient-derived cells revealed cell cycle abnormality. Our in vivo study using tedc1-/- zebrafish generated by CRISPR/Cas9 successfully recapitulated the growth impairment and cranial bone dysplasia as seen in our patients. The tedc1-/- mutant zebrafish were sterile and did not have developed gonads. Furthermore, we showed that biallelic TEDC1 deletion causes cilia abnormalities through defective acetylated tubulins.

    DOI: 10.1038/s41431-025-01802-3

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  • BET Inhibition Rescues Acinar-Ductal-Metaplasia and Ciliogenesis and Ameliorates Chronic Pancreatitis-Driven Changes in Mice With Loss of the Polarity Protein Par3.

    Shields MA, Metropulos AE, Spaulding C, Alzahrani KA, Hirose T, Ohno S, Pham TND, Munshi HG

    Cellular and molecular gastroenterology and hepatology   2024年8月

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    掲載種別:研究論文(学術雑誌)  

    <h4>Background & aims</h4>The apical-basal polarity of pancreatic acinar cells is essential for maintaining tissue architecture. However, the mechanisms by which polarity proteins regulate acinar pancreas injury and regeneration are poorly understood.<h4>Methods</h4>Cerulein-induced pancreatitis was induced in mice with conditional deletion of the polarity protein Par3 in the pancreas. The impact of Par3 loss on pancreas injury and regeneration was assessed by histologic analyses and transcriptional profiling by RNA sequencing. Mice were pretreated with the bromodomain and extraterminal domain (BET) inhibitor JQ1 before cotreatment with cerulein to determine the effect of BET inhibition on pancreas injury and regeneration.<h4>Results</h4>Initially, we show that Par3 is increased in acinar-ductal metaplasia (ADM) lesions present in human and mouse chronic pancreatitis specimens. Although Par3 loss disrupts tight junctions, Par3 is dispensable for pancreatogenesis. However, with aging, Par3 loss results in low-grade inflammation, acinar degeneration, and pancreatic lipomatosis. Par3 loss exacerbates acute pancreatitis-induced injury and chronic pancreatitis-induced acinar cell loss, promotes pancreatic lipomatosis, and prevents regeneration. Par3 loss also results in suppression of chronic pancreatitis-induced ADM and primary ciliogenesis. Notably, targeting BET proteins attenuates chronic pancreatitis-induced loss of primary cilia and promotes ADM in mice lacking pancreatic Par3. Targeting BET proteins also attenuates cerulein-induced acinar cell loss and enhances recovery of acinar cell mass and body weight of mice lacking pancreatic Par3.<h4>Conclusions</h4>Combined, this study demonstrates how Par3 restrains chronic pancreatitis-induced changes in the pancreas and identifies a potential role for BET inhibitors to attenuate pancreas injury and facilitate regeneration.

    DOI: 10.1016/j.jcmgh.2024.101389

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  • Chimera RNA transcribed from integrated HPV18 genome with adjacent host genomic region promotes oncogenic gene expression through condensate formation. 査読 国際誌

    Kazuki Furugori, Hidefumi Suzuki, Ryota Abe, Keiko Horiuchi, Tomohiko Akiyama, Tomonori Hirose, Atsushi Toyoda, Hidehisa Takahashi

    Genes to cells : devoted to molecular & cellular mechanisms   2024年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Most cervical cancers are caused by human papillomavirus (HPV) infection. In HeLa cells, the HPV18 viral genome is integrated at chromosome 8q24.21 and activates transcription of the proto-oncogene c-Myc. However, the mechanism of how the integrated HPV genome and its transcribed RNAs exhibit transcription activation function has not been fully elucidated. In this study, we found that HPV18 transcripts contain an enhancer RNA-like function to activate proximal genes including CCAT1-5L and c-Myc. We showed that the human genome-integrated HPV18 genes are activated by transcription coregulators including BRD4 and Mediator. The transcribed HPV18 RNAs form a liquid-like condensate at chromosome 8q24.21 locus, which in turn accumulates RNA polymerase II. Moreover, we focused on a relatively uncharacterized transcript from the upstream region of CCAT1, named URC. The URC RNA is transcribed as a chimera RNA with HPV18 and is composed of the 3'-untranslated region of the HPV18 transcript. We experimentally showed that the URC contributes to stabilization of HPV18 RNAs by supplying a polyadenylation site for the HPV18 transcript. Our findings suggest that integrated HPV18 at 8q24.21 locus produces HPV18-URC chimera RNA and promotes tumorigenesis through RNA-based condensate formation.

    DOI: 10.1111/gtc.13121

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  • The 3' Pol II pausing at replication-dependent histone genes is regulated by Mediator through Cajal bodies' association with histone locus bodies. 国際誌

    Hidefumi Suzuki, Ryota Abe, Miho Shimada, Tomonori Hirose, Hiroko Hirose, Keisuke Noguchi, Yoko Ike, Nanami Yasui, Kazuki Furugori, Yuki Yamaguchi, Atsushi Toyoda, Yutaka Suzuki, Tatsuro Yamamoto, Noriko Saitoh, Shigeo Sato, Chieri Tomomori-Sato, Ronald C Conaway, Joan W Conaway, Hidehisa Takahashi

    Nature communications   13 ( 1 )   2905 - 2905   2022年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Non-polyadenylated mRNAs of replication-dependent histones (RDHs) are synthesized by RNA polymerase II (Pol II) at histone locus bodies (HLBs). HLBs frequently associate with Cajal bodies (CBs), in which 3'-end processing factors for RDH genes are enriched; however, this association's role in transcription termination of RDH genes remains unclear. Here, we show that Pol II pauses immediately upstream of transcript end sites of RDH genes and Mediator plays a role in this Pol II pausing through CBs' association with HLBs. Disruption of the Mediator docking site for Little elongation complex (LEC)-Cap binding complex (CBC)-Negative elongation factor (NELF), components of CBs, interferes with CBs' association with HLBs and 3' Pol II pausing, resulting in increased aberrant unprocessed RDH gene transcripts. Our findings suggest Mediator's involvement in CBs' association with HLBs to facilitate 3' Pol II pausing and subsequent 3'-end processing of RDH genes by supplying 3'-end processing factors.

    DOI: 10.1038/s41467-022-30632-w

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  • Collagen XVII deficiency alters epidermal patterning. 国際誌

    Yunan Wang, Hiroyuki Kitahata, Hideyuki Kosumi, Mika Watanabe, Yu Fujimura, Shota Takashima, Shin-Ichi Osada, Tomonori Hirose, Wataru Nishie, Masaharu Nagayama, Hiroshi Shimizu, Ken Natsuga

    Laboratory investigation; a journal of technical methods and pathology   102 ( 6 )   581 - 588   2022年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Vertebrates exhibit patterned epidermis, exemplified by scales/interscales in mice tails and grooves/ridges on the human skin surface (microtopography). Although the role of spatiotemporal regulation of stem cells (SCs) has been implicated in this process, the mechanism underlying the development of such epidermal patterns is poorly understood. Here, we show that collagen XVII (COL17), a niche for epidermal SCs, helps stabilize epidermal patterns. Gene knockout and rescue experiments revealed that COL17 maintains the width of the murine tail scale epidermis independently of epidermal cell polarity. Skin regeneration after wounding was associated with slender scale epidermis, which was alleviated by overexpression of human COL17. COL17-negative skin in human junctional epidermolysis bullosa showed a distinct epidermal pattern from COL17-positive skin that resulted from revertant mosaicism. These results demonstrate that COL17 contributes to defining mouse tail scale shapes and human skin microtopography. Our study sheds light on the role of the SC niche in tissue pattern formation.

    DOI: 10.1038/s41374-022-00738-2

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  • De novo ATP1A3 variants cause polymicrogyria. 国際誌

    Satoko Miyatake, Mitsuhiro Kato, Takuma Kumamoto, Tomonori Hirose, Eriko Koshimizu, Takaaki Matsui, Hideyuki Takeuchi, Hiroshi Doi, Keisuke Hamada, Mitsuko Nakashima, Kazunori Sasaki, Akio Yamashita, Atsushi Takata, Kohei Hamanaka, Mai Satoh, Takabumi Miyama, Yuri Sonoda, Momoko Sasazuki, Hiroyuki Torisu, Toshiro Hara, Yasunari Sakai, Yushi Noguchi, Mazumi Miura, Yoko Nishimura, Kazuyuki Nakamura, Hideyuki Asai, Nodoka Hinokuma, Fuyuki Miya, Tatsuhiko Tsunoda, Masami Togawa, Yukihiro Ikeda, Nobusuke Kimura, Kaoru Amemiya, Asako Horino, Masataka Fukuoka, Hiroko Ikeda, Goni Merhav, Nina Ekhilevitch, Masaki Miura, Takeshi Mizuguchi, Noriko Miyake, Atsushi Suzuki, Shouichi Ohga, Hirotomo Saitsu, Hidehisa Takahashi, Fumiaki Tanaka, Kazuhiro Ogata, Chiaki Ohtaka-Maruyama, Naomichi Matsumoto

    Science advances   7 ( 13 )   2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Polymicrogyria is a common malformation of cortical development whose etiology remains elusive. We conducted whole-exome sequencing for 124 patients with polymicrogyria and identified de novo ATP1A3 variants in eight patients. Mutated ATP1A3 causes functional brain diseases, including alternating hemiplegia of childhood (AHC), rapid-onset dystonia parkinsonism (RDP), and cerebellar ataxia, areflexia, pes cavus, optic nerve atrophy, and sensorineural deafness (CAPOS). However, our patients showed no clinical features of AHC, RDP, or CAPOS and had a completely different phenotype: a severe form of polymicrogyria with epilepsy and developmental delay. Detected variants had different locations in ATP1A3 and different functional properties compared with AHC-, RDP-, or CAPOS-associated variants. In the developing cerebral cortex of mice, radial neuronal migration was impaired in neurons overexpressing the ATP1A3 variant of the most severe patients, suggesting that this variant is involved in cortical malformation pathogenesis. We propose a previously unidentified category of polymicrogyria associated with ATP1A3 abnormalities.

    DOI: 10.1126/sciadv.abd2368

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  • Type XVII collagen interacts with the aPKC-PAR complex and maintains epidermal cell polarity. 国際誌

    Mika Watanabe, Hideyuki Kosumi, Shin-Ichi Osada, Shota Takashima, Yunan Wang, Wataru Nishie, Tsukasa Oikawa, Tomonori Hirose, Hiroshi Shimizu, Ken Natsuga

    Experimental dermatology   30 ( 1 )   62 - 67   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Type XVII collagen (COL17) is a transmembrane protein expressed in the basal epidermis. COL17 serves as a niche for epidermal stem cells, and although its reduction has been implicated in altering cell polarity and ageing of the epidermis, it is unknown how COL17 affects epidermal cell polarity. Here, we uncovered COL17 as a binding partner of the aPKC-PAR complex, which is a key regulating factor of cell polarity. Immunoprecipitation-immunoblot assay and protein-protein binding assay revealed that COL17 interacts with aPKC and PAR3. COL17 deficiency or epidermis-specific aPKCλ deletion destabilized PAR3 distribution in the epidermis, while aPKCζ knockout did not. Asymmetrical cell division was pronounced in COL17-null neonatal paw epidermis. These results show that COL17 is pivotal for maintaining epidermal cell polarity. Our study highlights the previously unrecognized role of COL17 in the basal keratinocytes.

    DOI: 10.1111/exd.14196

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  • A phospho-switch controls RNF43-mediated degradation of Wnt receptors to suppress tumorigenesis

    Tadasuke Tsukiyama, Juqi Zou, Jihoon Kim, Shohei Ogamino, Yuki Shino, Takamasa Masuda, Alessandra Merenda, Masaki Matsumoto, Yoichiro Fujioka, Tomonori Hirose, Sayuri Terai, Hidehisa Takahashi, Tohru Ishitani, Keiichi I. Nakayama, Yusuke Ohba, Bon-Kyoung Koo, Shigetsugu Hatakeyama

    Nature Communications   11 ( 1 )   2020年12月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    <title>Abstract</title>
    Frequent mutation of the tumour suppressor RNF43 is observed in many cancers, particularly colon malignancies. RNF43, an E3 ubiquitin ligase, negatively regulates Wnt signalling by inducing degradation of the Wnt receptor Frizzled. In this study, we discover that RNF43 activity requires phosphorylation at a triplet of conserved serines. This phospho-regulation of RNF43 is required for zebrafish development and growth of mouse intestinal organoids. Cancer-associated mutations that abrogate RNF43 phosphorylation cooperate with active Ras to promote tumorigenesis by abolishing the inhibitory function of RNF43 in Wnt signalling while maintaining its inhibitory function in p53 signalling. Our data suggest that RNF43 mutations cooperate with KRAS mutations to promote multi-step tumorigenesis via the Wnt-Ras-p53 axis in human colon cancers. Lastly, phosphomimetic substitutions of the serine trio restored the tumour suppressive activity of extracellular oncogenic mutants. Therefore, harnessing phospho-regulation of RNF43 might be a potential therapeutic strategy for tumours with RNF43 mutations.

    DOI: 10.1038/s41467-020-18257-3

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    その他リンク: http://www.nature.com/articles/s41467-020-18257-3

  • Phosphorylation and dephosphorylation of Ser852 and Ser889 control the clustering, localization and function of PAR3. 国際誌

    Kazunari Yamashita, Keiko Mizuno, Kana Furukawa, Hiroko Hirose, Natsuki Sakurai, Maki Masuda-Hirata, Yoshiko Amano, Tomonori Hirose, Atsushi Suzuki, Shigeo Ohno

    Journal of cell science   133 ( 22 )   2020年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cell polarity is essential for various asymmetric cellular events, and the partitioning defective (PAR) protein PAR3 (encoded by PARD3 in mammals) plays a unique role as a cellular landmark to establish polarity. In epithelial cells, PAR3 localizes at the subapical border, such as the tight junction in vertebrates, and functions as an apical determinant. Although we know a great deal about the regulators of PAR3 localization, how PAR3 is concentrated and localized to a specific membrane domain remains an important question to be clarified. In this study, we demonstrate that ASPP2 (also known as TP53BP2), which controls PAR3 localization, links PAR3 and protein phosphatase 1 (PP1). The ASPP2-PP1 complex dephosphorylates a novel phosphorylation site, Ser852, of PAR3. Furthermore, Ser852- or Ser889-unphosphorylatable PAR3 mutants form protein clusters, and ectopically localize to the lateral membrane. Concomitance of clustering and ectopic localization suggests that PAR3 localization is a consequence of local clustering. We also demonstrate that unphosphorylatable forms of PAR3 exhibited a low molecular turnover and failed to coordinate rapid reconstruction of the tight junction, supporting that both the phosphorylated and dephosphorylated states are essential for the functional integrity of PAR3.

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  • The role of Mediator and Little Elongation Complex in transcription termination. 査読 国際誌

    Hidehisa Takahashi, Amol Ranjan, Shiyuan Chen, Hidefumi Suzuki, Mio Shibata, Tomonori Hirose, Hiroko Hirose, Kazunori Sasaki, Ryota Abe, Kai Chen, Yanfeng He, Ying Zhang, Ichigaku Takigawa, Tadasuke Tsukiyama, Masashi Watanabe, Satoshi Fujii, Midori Iida, Junichi Yamamoto, Yuki Yamaguchi, Yutaka Suzuki, Masaki Matsumoto, Keiichi I Nakayama, Michael P Washburn, Anita Saraf, Laurence Florens, Shigeo Sato, Chieri Tomomori-Sato, Ronald C Conaway, Joan W Conaway, Shigetsugu Hatakeyama

    Nature communications   11 ( 1 )   1063 - 1063   2020年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Mediator is a coregulatory complex that regulates transcription of Pol II-dependent genes. Previously, we showed that human Mediator subunit MED26 plays a role in the recruitment of Super Elongation Complex (SEC) or Little Elongation Complex (LEC) to regulate the expression of certain genes. MED26 plays a role in recruiting SEC to protein-coding genes including c-myc and LEC to small nuclear RNA (snRNA) genes. However, how MED26 engages SEC or LEC to regulate distinct genes is unclear. Here, we provide evidence that MED26 recruits LEC to modulate transcription termination of non-polyadenylated transcripts including snRNAs and mRNAs encoding replication-dependent histone (RDH) at Cajal bodies. Our findings indicate that LEC recruited by MED26 promotes efficient transcription termination by Pol II through interaction with CBC-ARS2 and NELF/DSIF, and promotes 3' end processing by enhancing recruitment of Integrator or Heat Labile Factor to snRNA or RDH genes, respectively.

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  • Gain-of-Function MN1 Truncation Variants Cause a Recognizable Syndrome with Craniofacial and Brain Abnormalities. 査読 国際誌

    Noriko Miyake, Hidehisa Takahashi, Kazuyuki Nakamura, Bertrand Isidor, Yoko Hiraki, Eriko Koshimizu, Masaaki Shiina, Kazunori Sasaki, Hidefumi Suzuki, Ryota Abe, Yayoi Kimura, Tomoko Akiyama, Shin-Ichi Tomizawa, Tomonori Hirose, Kohei Hamanaka, Satoko Miyatake, Satomi Mitsuhashi, Takeshi Mizuguchi, Atsushi Takata, Kazuyuki Obo, Mitsuhiro Kato, Kazuhiro Ogata, Naomichi Matsumoto

    American journal of human genetics   106 ( 1 )   13 - 25   2020年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    MN1 was originally identified as a tumor-suppressor gene. Knockout mouse studies have suggested that Mn1 is associated with craniofacial development. However, no MN1-related phenotypes have been established in humans. Here, we report on three individuals who have de novo MN1 variants that lead to a protein lacking the carboxyl (C) terminus and who presented with severe developmental delay, craniofacial abnormalities with specific facial features, and structural abnormalities in the brain. An in vitro study revealed that the deletion of the C-terminal region led to increased protein stability, an inhibitory effect on cell proliferation, and enhanced MN1 aggregation in nuclei compared to what occurred in the wild type, suggesting that a gain-of-function mechanism is involved in this disease. Considering that C-terminal deletion increases the fraction of intrinsically disordered regions of MN1, it is possible that altered phase separation could be involved in the mechanism underlying the disease. Our data indicate that MN1 participates in transcriptional regulation of target genes through interaction with the transcription factors PBX1, PKNOX1, and ZBTB24 and that mutant MN1 impairs the binding with ZBTB24 and RING1, which is an E3 ubiquitin ligase. On the basis of our findings, we propose the model that C-terminal deletion interferes with MN1's interaction molecules related to the ubiquitin-mediated proteasome pathway, including RING1, and increases the amount of the mutant protein; this increase leads to the dysregulation of MN1 target genes by inhibiting rapid MN1 protein turnover.

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  • Atypical protein kinase C isoforms differentially regulate directional keratinocyte migration during wound healing 査読

    Noguchi, N., Hirose, T., Suzuki, T., Kagaya, M., Chida, K., Ohno, S., Manabe, M., Osada, S.-I.

    Journal of Dermatological Science   93 ( 2 )   101 - 108   2019年2月

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    掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.jdermsci.2019.01.001

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  • aPKC controls endothelial growth by modulating c-Myc via FoxO1 DNA-binding ability. 査読 国際誌

    Meghan Riddell, Akiko Nakayama, Takao Hikita, Fatemeh Mirzapourshafiyi, Takuji Kawamura, Ayesha Pasha, Mengnan Li, Mikio Masuzawa, Mario Looso, Tim Steinbacher, Klaus Ebnet, Michael Potente, Tomonori Hirose, Shigeo Ohno, Ingrid Fleming, Stefan Gattenlöhner, Phyu P Aung, Thuy Phung, Osamu Yamasaki, Teruki Yanagi, Hiroshi Umemura, Masanori Nakayama

    Nature communications   9 ( 1 )   5357 - 5357   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Strict regulation of proliferation is vital for development, whereas unregulated cell proliferation is a fundamental characteristic of cancer. The polarity protein atypical protein kinase C lambda/iota (aPKCλ) is associated with cell proliferation through unknown mechanisms. In endothelial cells, suppression of aPKCλ impairs proliferation despite hyperactivated mitogenic signaling. Here we show that aPKCλ phosphorylates the DNA binding domain of forkhead box O1 (FoxO1) transcription factor, a gatekeeper of endothelial growth. Although mitogenic signaling excludes FoxO1 from the nucleus, consequently increasing c-Myc abundance and proliferation, aPKCλ controls c-Myc expression via FoxO1/miR-34c signaling without affecting its localization. We find this pathway is strongly activated in the malignant vascular sarcoma, angiosarcoma, and aPKC inhibition reduces c-Myc expression and proliferation of angiosarcoma cells. Moreover, FoxO1 phosphorylation at Ser218 and aPKC expression correlates with poor patient prognosis. Our findings may provide a potential therapeutic strategy for treatment of malignant cancers, like angiosarcoma.

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  • PAR-3 controls endothelial planar polarity and vascular inflammation under laminar flow. 査読 国際誌

    Hikita T, Mirzapourshafiyi F, Barbacena P, Riddell M, Pasha A, Li M, Kawamura T, Brandes RP, Hirose T, Ohno S, Gerhardt H, Matsuda M, Franco CA, Nakayama M

    EMBO reports   19 ( 9 )   2018年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.15252/embr.201745253

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  • Biallelic Mutations in Nuclear Pore Complex Subunit NUP107 Cause Early-Childhood-Onset Steroid-Resistant Nephrotic Syndrome 査読

    Noriko Miyake, Hiroyasu Tsukaguchi, Eriko Koshimizu, Akemi Shono, Satoko Matsunaga, Masaaki Shiina, Yasuhiro Mimura, Shintaro Imamura, Tomonori Hirose, Koji Okudela, Kandai Nozu, Yuko Akioka, Motoshi Hattori, Norishige Yoshikawa, Akiko Kitamura, Hae Il Cheong, Shoji Kagami, Michiaki Yamashita, Atsushi Fujita, Satoko Miyatake, Yoshinori Tsurusaki, Mitsuko Nakashima, Hirotomo Saitsu, Kenichi. Ohashi, Naoko Imamoto, Akihide Ryo, Kazuhiro Ogata, Kazumoto Iijima, Naomichi Matsumoto

    AMERICAN JOURNAL OF HUMAN GENETICS   97 ( 4 )   555 - 566   2015年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    The nuclear pore complex (NPC) is a huge protein complex embedded in the nuclear envelope. It has central functions in nucleocytoplasmic transport, nuclear framework, and gene regulation. Nucleoporin 107 kDa (NUP107) is a component of the NPC central scaffold and is an essential protein in all eukaryotic cells. Here, we report on biallelic NUP107 mutations in nine affected individuals who are from five unrelated families and show early-onset steroid-resistant nephrotic syndrome (SRN). These individuals have pathologically focal segmental glomerulosclerosis, a condition that leads to end-stage renal disease with high frequency. NUP107 is ubiquitously expressed, including in glomerular podocytes. Three of four NUP107 mutations detected in the affected individuals hamper NUP107 binding to NUP133 (nucleoporin 133 kDa) and NUP107 incorporation into NPCs in vitro. Zebrafish with nup107 knockdown generated by morpholino oligonucleotides displayed hypoplastic glomerulus structures and abnormal podocyte foot processes, thereby mimicking the pathological changes seen in the kidneys of the SRNS individuals with NUP107 mutations. Considering the unique properties of the podocyte (highly differentiated foot-process architecture and slit membrane and the inability to regenerate), we propose a "podocyte-injury model" as the pathomechanism for SRNS due to biallelic NUP107 mutations.

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  • APKCλ maintains the integrity of the glomerular slit diaphragm through trafficking of nephrin to the cell surface 査読

    Daisuke Satoh, Tomonori Hirose, Yutaka Harita, Yutaka Harita, Chikara Daimon, Tomonori Harada, Hidetake Kurihara, Akio Yamashita, Shigeo Ohno, Shigeo Ohno

    Journal of Biochemistry   156 ( 2 )   115 - 128   2014年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The slit diaphragm (SD), the specialized intercellular junction between renal glomerular epithelial cells (podocytes), provides a selective-filtration barrier in renal glomeruli. Dysfunction of the SD results in glomerular diseases that are characterized by disappearance of SD components, such as nephrin, from the cell surface. Although the importance of endocytosis and degradation of SD components for the maintenance of SD integrity has been suggested, the dynamic nature of the turnover of intact cell-surface SD components remained unclear. Using isolated rat glomeruli we show that the turnover rates of cell-surface SD components are relatively high; they almost completely disappear from the cell surface within minutes. The exocytosis, but not endocytosis, of heterologously expressed nephrin requires the kinase activity of the cell polarity regulator atypical protein kinase C (aPKC). Consistently, we demonstrate that podocyte-specific deletion of aPKCλ resulted in a decrease of cell-surface localization of SD components, causing massive proteinuria. In conclusion, the regulation of SD turnover by aPKC is crucial for the maintenance of SD integrity and defects in aPKC signalling can lead to proteinuria. These findings not only reveal the pivotal importance of the dynamic turnover of cell-surface SD components but also suggest a novel pathophysiological basis in glomerular disease. © 2014 The Authors 2014. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

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  • Loss of aPKCλ in differentiated neurons disrupts the polarity complex but does not induce obvious neuronal loss or disorientation in mouse brains. 査読

    Yamanaka T, Tosaki A, Kurosawa M, Akimoto K, Hirose T, Ohno S, Hattori N, Nukina N

    PloS one   8 ( 12 )   e84036   2013年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:12  

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  • Spatial regulation of VEGF receptor endocytosis in angiogenesis 査読

    Masanori Nakayama, Akiko Nakayama, Max van Lessen, Hiroyuki Yamamoto, Sarah Hoffmann, Hannes C. A. Drexler, Norimichi Itoh, Tomonori Hirose, Georg Breier, Dietmar Vestweber, Jonathan A. Cooper, Shigeo Ohno, Kozo Kaibuchi, Ralf H. Adams

    NATURE CELL BIOLOGY   15 ( 3 )   249 - 260   2013年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Activities as diverse as migration, proliferation and patterning occur simultaneously and in a coordinated fashion during tissue morphogenesis. In the growing vasculature, the formation of motile, invasive and filopodia-carrying endothelial sprouts is balanced with the stabilization of blood-transporting vessels. Here, we show that sprouting endothelial cells in the retina have high rates of VEGF uptake, VEGF receptor endocytosis and turnover. These internalization processes are opposed by atypical protein kinase C activity in more stable and mature vessels. aPKC phosphorylates Dab2, a clathrin-associated sorting protein that, together with the transmembrane protein ephrin-B2 and the cell polarity regulator PAR-3, enables VEGF receptor endocytosis and downstream signal transduction. Accordingly, VEGF receptor internalization and the angiogenic growth of vascular beds are defective in loss-of-function mice lacking key components of this regulatory pathway. Our work uncovers how vessel growth is dynamically controlled by local VEGF receptor endocytosis and the activity of cell polarity proteins.

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  • Tumor Type-Dependent Function of the Par3 Polarity Protein in Skin Tumorigenesis 査読

    Sandra Iden, Wilhelmina E. van Riel, Ronny Schafer, Ji-Ying Song, Tomonori Hirose, Shigeo Ohno, John G. Collard

    CANCER CELL   22 ( 3 )   389 - 403   2012年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    Cell polarization is crucial during development and tissue homeostasis and is regulated by conserved proteins of the Scribble, Crumbs, and Par complexes. In mouse skin tumorigenesis, Par3 deficiency results in reduced papilloma formation and growth. Par3 mediates its tumor-promoting activity through regulation of growth and survival, since Par3 deletion increases apoptosis and reduces growth in vivo and in vitro. In contrast, Par3-deficient mice are predisposed to formation of keratoacanthomas, cutaneous tumors thought to originate from different cellular origin and frequently observed in humans. Par3 expression is reduced in both mouse and human keratoacanthomas, indicating tumor-suppressive properties of Par3. Our results identify a dual function of Par3 in skin cancer, with both pro-oncogenic and tumor-suppressive activity depending on the tumor type.

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  • Prepubertal angiotensin blockade exerts long-term therapeutic effect through sustained ATRAP activation in salt-sensitive hypertensive rats 査読

    Toru Dejima, Kouichi Tamura, Hiromichi Wakui, Akinobu Maeda, Masato Ohsawa, Tomohiko Kanaoka, Sona Haku, Azushima Kengo, Shin-ichiro Masuda, Atsu-ichiro Shigenaga, Koichi Azuma, Miyuki Matsuda, Machiko Yabana, Tomonori Hirose, Kazuaki Uchino, Kazuo Kimura, Yoji Nagashima, Satoshi Umemura

    JOURNAL OF HYPERTENSION   29 ( 10 )   1919 - 1929   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:LIPPINCOTT WILLIAMS & WILKINS  

    Objective We previously showed that the molecule interacting with Ang II type 1 receptor (AT1R), ATRAP, promotes AT1R internalization and attenuates AT1R-mediated pathological responses. In this study we examined whether the regulation of renal ATRAP expression is related to the development of salt-induced hypertension and renal injury as well as to the beneficial effects of AT1R blockade.
    Methods and results Dahl Iwai salt-sensitive hypertensive and Dahl Iwai salt-resistant rats were divided into six groups for the administration of vehicle or olmesartan either continuously from 3 to 16 weeks, or transiently from weaning to puberty (3-10 weeks), and fed high salt diet from 6 to 16 weeks. In Dahl Iwai salt-sensitive rats, not only continuous, but also prepubertal olmesartan treatment improved hypertension at 15 weeks. Renal ATRAP expression was suppressed in vehicle-treated Dahl Iwai salt-sensitive rats, concomitant with up-regulation of renal oxidative stress, inflammation and fibrosis-related markers such as p22phox, TGF-beta, fibronectin, monocyte chemotactic protein-1 and type 1 collagen. However, prepubertal as well as continuous olmesartan treatment recovered the suppressed renal ATRAP expression and inhibited the renal activation of p22phox, TGF-beta, fibronectin, MCP-1 and type 1 collagen. In Dahl Iwai salt-resistant rats, such suppression of renal ATRAP expression or induction of renal pathological responses by salt loading was not observed.
    Conclusions These results indicate that prepubertal transient blockade of AT1R signaling exerts a long-term therapeutic effect on salt-induced hypertension and renal injury in Dahl Iwai salt-sensitive rats, partly through a sustained enhancement of renal ATRAP expression, thereby suggesting ATRAP a novel molecular target in salt-induced hypertension and renal injury. J Hypertens 29:1919-1929 (C) 2011 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

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  • Involvement of Runx3 in the basal transcriptional activation of the mouse angiotensin II type 1 receptor-associated protein gene 査読

    Miyuki Matsuda, Kouichi Tamura, Hiromichi Wakui, Toru Dejima, Akinobu Maeda, Masato Ohsawa, Tomohiko Kanaoka, Sona Haku, Kengo Azushima, Hiroko Yamasaki, Daisuke Saito, Tomonori Hirose, Yohei Maeshima, Yoji Nagashima, Satoshi Umemura

    PHYSIOLOGICAL GENOMICS   43 ( 14 )   884 - 894   2011年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER PHYSIOLOGICAL SOC  

    Matsuda M, Tamura K, Wakui H, Dejima T, Maeda A, Ohsawa M, Kanaoka T, Haku S, Azushima K, Yamasaki H, Saito D, Hirose T, Maeshima Y, Nagashima Y, Umemura S. Involvement of Runx3 in the basal transcriptional activation of the mouse angiotensin II type 1 receptor-associated protein gene. Physiol Genomics 43: 884-894, 2011. First published May 17, 2011; doi:10.1152/physiolgenomics.00005.2011.-We previously cloned a molecule that interacts with angiotensin II type 1 (AT1) receptor to exert an inhibitory function on AT1 receptor signaling that we named ATRAP/Agtrap (for AT1 receptor-associated protein). In the present study we examined the regulation of basal ATRAP gene expression using renal distal convoluted tubule cells. We found that serum starvation upregulated basal expression of ATRAP gene, a response that required de novo mRNA and protein synthesis. Luciferase assay revealed that the proximal promoter region directs transcription and that a putative binding site of runt-related transcription factors (RBE) is important for transcriptional activation. The results of RBE-decoy transfection and endogenous knockdown by small interference RNA showed that the runt-related transcription factor Runx3 is involved in ATRAP gene expression. Chromatin immunoprecipitation assay also supported the binding of Runx3 to the ATRAP promoter in renal distal convoluted tubule cells. Immunohistochemistry demonstrated the expression of Runx3 and ATRAP proteins in the distal convoluted and connecting tubules of the kidney in consecutive sections. Furthermore, the Runx3 immunostaining was decreased together with a concomitant suppression of ATRAP expression in the affected kidney after 7 days of unilateral ureteral obstruction. These findings indicate that Runx3 plays a role in ATRAP gene expression in renal distal tubular cells both in vitro and in vivo.

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  • Intrarenal suppression of angiotensin II type 1 receptor binding molecule in angiotensin II-infused mice 査読

    Hiromichi Wakui, Kouichi Tamura, Miyuki Matsuda, Yunzhe Bai, Toru Dejima, Atsu-ichiro Shigenaga, Shin-ichiro Masuda, Koichi Azuma, Akinobu Maeda, Tomonori Hirose, Tomoaki Ishigami, Yoshiyuki Toya, Machiko Yabana, Susumu Minamisawa, Satoshi Umemura

    AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY   299 ( 5 )   F991 - F1003   2010年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER PHYSIOLOGICAL SOC  

    Wakui H, Tamura K, Matsuda M, Bai Y, Dejima T, Shigenaga A, Masuda S, Azuma K, Maeda A, Hirose T, Ishigami T, Toya Y, Yabana M, Minamisawa S, Umemura S. Intrarenal suppression of angiotensin II type 1 receptor binding molecule in angiotensin II-infused mice. Am J Physiol Renal Physiol 299: F991-F1003, 2010. First published August 25, 2010; doi:10.1152/ajprenal.00738.2009.-ATRAP [ANG II type 1 receptor (AT1R)-associated protein] is a molecule which directly interacts with AT1R and inhibits AT1R signaling. The aim of this study was to examine the effects of continuous ANG II infusion on the intrarenal expression and distribution of ATRAP and to determine the role of AT1R signaling in mediating these effects. C57BL/6 male mice were subjected to vehicle or ANG II infusions at doses of 200, 1,000, or 2,500 ng . kg(-1) . min(-1) for 14 days. ANG II infusion caused significant suppression of ATRAP expression in the kidney but did not affect ATRAP expression in the testis or liver. Although only the highest ANG II dose (2,500 ng . kg(-1) . min(-1)) provoked renal pathological responses, such as an increase in the mRNA expression of angiotensinogen and the alpha-subunit of the epithelial sodium channel, ANG II-induced decreases in ATRAP were observed even at the lowest dose (200 ng . kg(-1) . min(-1)), particularly in the outer medulla of the kidney, based on immunohistochemical staining and Western blot analysis. The decrease in renal ATRAP expression by ANG II infusion was prevented by treatment with the AT1R-specific blocker olmesartan. In addition, the ANG II-mediated decrease in renal ATRAP expression through AT1R signaling occurred without an ANG II-induced decrease in plasma membrane AT1R expression in the kidney. On the other hand, a transgenic model increase in renal ATRAP expression beyond baseline was accompanied by a constitutive reduction of renal plasma membrane AT1R expression and by the promotion of renal AT1R internalization as well as the decreased induction of angiotensinogen gene expression in response to ANG II. These results suggest that the plasma membrane AT1R level in the kidney is modulated by intrarenal ATRAP expression under physiological and pathophysiological conditions in vivo.

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  • Intrarenal suppression of angiotensin II type 1 receptor binding molecule in angiotensin II-infused mice 査読

    Hiromichi Wakui, Kouichi Tamura, Miyuki Matsuda, Yunzhe Bai, Toru Dejima, Atsu-Ichiro Shigenaga, Shin-Ichiro Masuda, Koichi Azuma, Akinobu Maeda, Tomonori Hirose, Tomoaki Ishigami, Yoshiyuki Toya, Machiko Yabana, Susumu Minamisawa, Satoshi Umemura

    American Journal of Physiology - Renal Physiology   299 ( 5 )   F991 - F1003   2010年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    ATRAP [ANG II type 1 receptor (AT1R)-associated protein] is a molecule which directly interacts with AT1R and inhibits AT1R signaling. The aim of this study was to examine the effects of continuous ANG II infusion on the intrarenal expression and distribution of ATRAP and to determine the role of AT1R signaling in mediating these effects. C57BL/6 male mice were subjected to vehicle or ANG II infusions at doses of 200, 1,000, or 2,500 ng·kg -1·min-1 for 14 days. ANG II infusion caused significant suppression of ATRAP expression in the kidney but did not affect ATRAP expression in the testis or liver. Although only the highest ANG II dose (2,500 ng·kg-1·min-1) provoked renal pathological responses, such as an increase in the mRNA expression of angiotensinogen and the α-subunit of the epithelial sodium channel, ANG II-induced decreases in ATRAP were observed even at the lowest dose (200 ng·kg-1·min-1), particularly in the outer medulla of the kidney, based on immunohistochemical staining and Western blot analysis. The decrease in renal ATRAP expression by ANG II infusion was prevented by treatment with the AT1R-specific blocker olmesartan. In addition, the ANG II-mediated decrease in renal ATRAP expression through AT1R signaling occurred without an ANG II-induced decrease in plasma membrane AT1R expression in the kidney. On the other hand, a transgenic model increase in renal ATRAP expression beyond baseline was accompanied by a constitutive reduction of renal plasma membrane AT1R expression and by the promotion of renal AT1R internalization as well as the decreased induction of angiotensinogen gene expression in response to ANG II. These results suggest that the plasma membrane AT1R level in the kidney is modulated by intrarenal ATRAP expression under physiological and pathophysiological conditions in vivo. Copyright © 2010 the American Physiological Society.

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  • Expression of angiotensin II type 1 receptor-interacting molecule in normal human kidney and IgA nephropathy 査読

    Shin-ichiro Masuda, Kouichi Tamura, Hiromichi Wakui, Akinobu Maeda, Toru Dejima, Tomonori Hirose, Masao Toyoda, Koichi Azuma, Masato Ohsawa, Tomohiko Kanaoka, Mai Yanagi, Shin-ichiro Yoshida, Hiroshi Mitsuhashi, Miyuki Matsuda, Tomoaki Ishigami, Yoshiyuki Toya, Daisuke Suzuki, Yoji Nagashima, Satoshi Umemura

    AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY   299 ( 4 )   F720 - F731   2010年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER PHYSIOLOGICAL SOC  

    Masuda S, Tamura K, Wakui H, Maeda A, Dejima T, Hirose T, Toyoda M, Azuma K, Ohsawa M, Kanaoka T, Yanagi M, Yoshida S, Mitsuhashi H, Matsuda M, Ishigami T, Toya Y, Suzuki D, Nagashima Y, Umemura S. Expression of angiotensin II type 1 receptor-interacting molecule in normal human kidney and IgA nephropathy. Am J Physiol Renal Physiol 299: F720-F731, 2010. First published August 4, 2010; doi:10.1152/ajprenal.00667.2009.-The intrarenal renin-angiotensin system plays a crucial role in the regulation of renal circulation and sodium reabsorption through the activation of vascular, glomerular, and tubular angiotensin II type 1 (AT(1)) receptor signaling. We previously cloned a molecule that specifically interacted with the murine AT(1) receptor to inhibit AT(1) receptor signaling, which we named ATRAP (for AT(1) receptor-associated protein). Since murine ATRAP was shown to be highly expressed in the kidney, in the present study we investigated expression and distribution of human ATRAP in normal kidney and renal biopsy specimens from patients with IgA nephropathy. In the normal human kidney, both ATRAP mRNA and protein were widely and abundantly distributed along the renal tubules from Bowman&apos;s capsule to the medullary collecting ducts. In all renal tubular epithelial cells, the ATRAP protein colocalized with the AT(1) receptor. In renal biopsy specimens with IgA nephropathy, a significant positive correlation between ATRAP and AT(1) receptor gene expression was observed. There was also a positive relationship between tubulointerstitial ATRAP expression and the estimated glomerular filtration rate in patients with IgA nephropathy. Furthermore, we examined the function of the tubular AT(1) receptor using an immortalized cell line of mouse distal convoluted tubule cells (mDCT) and found that overexpression of ATRAP by adenoviral gene transfer suppressed the angiotensin II-mediated increases in transforming growth factor-beta production in mDCT cells. These findings suggest that ATRAP might play a role in balancing the renal renin-angiotensin system synergistically with the AT(1) receptor by counterregulatory effects in IgA nephropathy and propose an antagonistic effect of tubular ATRAP on AT(1) receptor signaling.

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  • Expression of Toll-like receptor 9 in renal podocytes in childhood-onset active and inactive lupus nephritis 査読

    Hiroyuki Machida, Shuichi Ito, Tomonori Hirose, Fumihiko Takeshita, Hisashi Oshiro, Tomoko Nakamura, Masaaki Mori, Yoshiaki Inayama, Kunimasa Yan, Naoto Kobayashi, Shumpei Yokota

    NEPHROLOGY DIALYSIS TRANSPLANTATION   25 ( 8 )   2530 - 2537   2010年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Background. Childhood-onset systemic lupus erythematosus (SLE) is frequently complicated with lupus nephritis (LN), which is characterized by the deposition of DNA-containing immune complex to the glomerulus. Toll-like receptor 9 (TLR9), capable of recognizing the microbially derived CpG oligonucleotide, plays a crucial role in the innate immunity. TLR9 is also assumed to be related to the aetiology of SLE in the recognition of anti-DNA antibody-containing immune complex, but this remains controversial. We conducted a study to elucidate the association between TLR9 and LN in childhood-onset SLE.
    Methods. We compared the expression and localization of TLR9 and the slit membrane-related protein in the biopsied kidney sample by immunostaining in four children with active or inactive LN. We also evaluated their laboratory findings, such as anti-DNA antibody, complement and proteinuria at biopsy, to assess the correlation to the findings of the immunostaining. Results. TLR9 is not expressed in a normal control kidney. However, TLR9 develops in podocytes only in active LN but disappears in remission. Meanwhile, the slit membrane-related proteins such as nephrin, podocin and synaptopodin in podocytes express clearly and uniformly in remission, but their expression is markedly diminished in active LN, which results in podocyte injury. When TLR9 is expressed in podocytes, all the patients simultaneously showed hypocomplementaemia, high titre of anti-double-stranded DNA (dsDNA) antibody and proteinuria.
    Conclusion. Injured podocytes in active LN express TLR9. This expression could be associated with proteinuria and increased anti-dsDNA antibody. This is the first report indicating that TLR9 is involved in the aetiology of LN and that it may play some role in podocyte injury.

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  • ASPP2 Regulates Epithelial Cell Polarity through the PAR Complex 査読

    Weili Cong, Tomonori Hirose, Yutaka Harita, Akio Yamashita, Keiko Mizuno, Hisashi Hirano, Shigeo Ohno

    CURRENT BIOLOGY   20 ( 15 )   1408 - 1414   2010年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    The PAR complex, consisting of the evolutionarily conserved PAR-3, PAR-6, and aPKC, regulates cell polarity in many cell types, including epithelial cells [1-4]. Consistently, genetic manipulation of its components affects tissue integrity in multiple biological systems [5-9]. However, the regulatory mechanisms of the PAR complex remain obscure. We report here that apoptosis-stimulating protein of p53 (ASPP2 or TP53BP2), which binds to the tumor suppressor p53 and stimulates its proapoptotic function [10-12], positively regulates epithelial cell polarity by associating with the PAR complex. ASPP2 interacts and colocalizes with PAR-3 at apical cell-cell junctions in the polarized epithelial cells. Depletion of ASPP2 in epithelial cells causes defects in cell polarity, such as the formation of tight junctions and the maintenance and development of apical membrane domains. Moreover, depletion of ASPP2 causes a defect in PAR-3 localization, as well as vice versa. Furthermore, disturbance in the interaction between ASPP2 and PAR-3 causes defects in cell polarity. We conclude that ASPP2 regulates epithelial cell polarity in cooperation with PAR-3 to form an active PAR complex. Our results, taken together with the known functions of ASPP2, suggest a close relationship between cell polarity and other cell regulatory mechanisms mediated by ASPP2.

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  • aPKC restricts the basolateral determinant PtdIns(3,4,5)P3 to the basal region 査読

    Shoukichi Takahama, Tomonori Hirose, Shigeo Ohno

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   368 ( 2 )   249 - 255   2008年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Both PtdIns(3,4,5)P3 (PIP3) and atypical protein kinase C (aPKC) play central roles in the polarization of many cell types. In epithelial cells, both PIP3 and aPKC are required for the development of apico-basolateral membrane polarity. However, the relationship between PIP3 and aPKC during the establishment and maintenance of polarized membrane domains remains to be clarified. We show that depolarized MDCK cells retain a polarized basal distribution of PIP3, supporting a role for PIP3 in determining the basolateral membrane domain. Importantly, overexpression of a kinase-negative mutant of aPKC lambda (aPKC lambda kn) impaired the basal distribution of PIP3, indicating that aPKC kinase activity is required for the restriction of PIP3 to the basal region. In support of this, overexpression of aPKC lambda kn during polarization, but not after polarization, caused whole membrane distribution of PIP3 as well as defects in epithelial polarization. (C) 2008 Elsevier Inc. All rights reserved.

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  • Loss of partitioning-defective-3/isotype-specific interacting protein (Par-3/ASIP) in the elongating spermatid of RA175 (IGSF4A/SynCAM)-deficient mice 査読

    Eriko Fujita, Yuko Tanabe, Tomonori Hirose, Michel Aurrand-Lions, Tadashi Kasahara, Beat A. Imhof, Shigeo Ohno, Takashi Momoi

    AMERICAN JOURNAL OF PATHOLOGY   171 ( 6 )   1800 - 1810   2007年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC INVESTIGATIVE PATHOLOGY, INC  

    IGSF4a/RA175/SynCAM (RA175) and junctional adhesion molecules (Jams) are members of the immunoglobulin superfamily with a PDZ-binding domain at their C termini. Deficiency of Ra175 (Ra175(-/-)) as well as jam-C deficiency (Jam-C-/-) causes the defect of the spermatid differentiation, oligo-astheno-teratozoospermia. Ra175(-/-) elongating spermatids; fail to mature further, whereas Jam-C-/- round spermatids lose cell polarity, and most of Jam-C-/- elongated spermatids are completely lost. RA175 and Jam-C seem to have similar but distinct functional roles during spermatid. differentiation. Here we show that the cell polarity protein Par-3 with PDZ domains, a binding partner of jams, is one of the associated proteins of the cytoplasmic region of RA175 in testis. Par-3 and Jam-C are partly co-localized with RA175 in the elongating and elongated spermatids; their distributions overlapped with that of RA175 on the tips of the dorsal region of the head of the elongating spermatid (steps 9 to 12) in the wild type. In the Ra175(-/-)-elongating spermatid, Par-3 was absent, and Jam-C was absent or abnormally localized. The RA175 formed a ternary complex with Jam-C via interaction with Par-3. The lack of the ternary complex in the Ra175(-/-) elongating spermatid may cause the defect of the specialized adhesion structures, resulting in the oligo-astheno-teratozoospermia.

    DOI: 10.2353/ajpath.2007.070261

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  • Neonatal pancreatic cells redifferentiate into both neural and pancreatic lineages 査読

    Wenping Zhao, Tomonori Hirose, Momotaro Ishikawa, Yuji Oshima, Syu-Ichi Hirai, Shigeo Ohno, Hideki Taniguchi

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   352 ( 1 )   84 - 90   2007年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Studies of islet neogenesis have suggested that the regeneration of islet cells can be achieved through redifferentiation of pre-existing islet cells. However, this hypothesis is largely unproven and fails to account for the diversity of observed islet neogenesis. Here we show that cultured neonatal pancreatic cells dedifferentiate into beta III tubulin-expressing precursors, which then expand and redifferentiate into both neural and pancreatic lineage progenies. Redifferentiation happens not only in the islet cells, but also in the ductal cells that may represent what are called ductal origin "pancreatic stem cells". The it? vitro redifferentiation of neonatal pancreatic cells recapitulates the embryonic development by sequential endocrine differentiation accompanied by the coexpression of neuronal marker beta III tubulin with endocrine hormones until terminal differentiation. The neuronal differentiation of pancreatic cells, however, occurs prior to endocrine differentiation and gradually becomes dominant, thus implying a default neuronal lineage specification for cultured pancreatic cells. (c) 2006 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2006.10.179

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  • sPAR-3, a splicing variant of PAR-3, shows cellular localization and an expression pattern different from that of PAR-3 during enterocyte polarization 査読

    T Yoshii, K Mizuno, T Hirose, A Nakajima, H Sekihara, S Ohno

    AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY   288 ( 3 )   G564 - G570   2005年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER PHYSIOLOGICAL SOC  

    PAR-3 (partitioning-defective) is a scaffold-like PDZ (postsynaptic density-95/discs large/zonula occludens-1) domain-containing protein that forms a complex with PAR-6 and atypical PKC, localizes to tight junctions, and contributes to the formation of functional tight junctions. There are several alternatively spliced isoforms of PAR-3, although their physiological significance remains unknown. In this study, we show that one of the major isoforms of PAR-3, sPAR-3, is predominantly expressed in the Caco-2 cells derived from colon carcinoma and is used as a model to investigate the events involved in the epithelial cell differentiation and cell polarity development. During the polarization of Caco-2 cells, the expression of PAR-3 increases as do those of other cell-cell junction proteins, whereas the expression of sPAR-3 decreases. Biochemical characterization revealed that sPAR-3 associates with atypical PKC, as does PAR-3. On the other hand, immunofluorescence microscopy revealed that sPAR-3 does not concentrate at the cell-cell contact region in fully polarized cells, whereas it concentrates at premature cell-cell junctions. This makes a contrast to PAR-3, which concentrates at tight junctions in fully polarized cells. These results provide evidence suggesting the difference in the role between sPAR-3 and PAR-3 in epithelial cells.

    DOI: 10.1152/ajpgi.00426.2003

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  • Self-association of PAR-3-mediated by the conserved N-terminal domain contributes to the development of epithelial tight junctions 査読

    K Mizuno, A Suzuki, T Hirose, K Kitamura, K Kutsuzawa, M Futaki, Y Amano, S Ohno

    JOURNAL OF BIOLOGICAL CHEMISTRY   278 ( 33 )   31240 - 31250   2003年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    PAR-3 is a scaffold-like PDZ-containing protein that forms a complex with PAR-6 and atypical protein kinase C (PAR-3-atypical protein kinase C-PAR-6 complex) and contributes to the establishment of cell polarity in a wide variety of biological contexts. In mammalian epithelial cells, it localizes to tight junctions, the most apical end of epithelial cell-cell junctions, and contributes to the formation of functional tight junctions. However, the mechanism by which PAR-3 localizes to tight junctions and contributes to their formation remains to be clarified. Here we show that the N-terminal conserved region, CR1-(1-86), and the sequence 937-1,024 are required for its recruitment to the most apical side of the cell-cell contact region in epithelial Madin-Darby canine kidney cells. We also show that CR1 self-associates to form an oligomeric complex in vivo and in vitro. Further, overexpression of CR1 in Madin-Darby canine kidney cells disturbs the distribution of atypical protein kinase C and PAR-6 as well as PAR-3 and delays the formation of functional tight junctions. These results support the notion that the CR1- mediated self-association of the PAR-3-containing protein complex plays a role during the formation of functional tight junctions.

    DOI: 10.1074/jbc.M303593200

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  • Mammalian LgI forms a protein complex with PAR-6 and aPKC independently of PAR-3 to regulate epithelial cell polarity 査読

    T Yamanaka, Y Horikoshi, Y Sugiyama, C Ishiyama, A Suzuki, T Hirose, A Iwamatsu, A Shinohara, S Ohno

    CURRENT BIOLOGY   13 ( 9 )   734 - 743   2003年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CELL PRESS  

    Background: Epithelial cells have apicobasal polarity and an asymmetric junctional complex that provides the bases for development and tissue maintenance. In both vertebrates and invertebrates, the evolutionarily conserved protein complex, PAR-6/aPKC/PAR-3, localizes to the subapical region and plays critical roles in the establishment of a junctional complex and cell polarity. In Drosophila, another set of proteins called tumor suppressors, such as LgI, which localize separately to the basolateral membrane domain but genetically interact with the subapical proteins, also contribute to the establishment of cell polarity. However, how physically separated proteins interact remains to be clarified.
    Results: We show that mammalian LgI competes for PAR-3 in forming an independent complex with PAR-6/ aPKC. During cell polarization, mLgI initially colocalizes with PAR-6/aPKC at the cell-cell contact region and is phosphorylated by aPKC, followed by segregation from apical PAR-6/aPKC to the basolateral membrane after cells are polarized. Overexpression studies establish that increased amounts of the mLgI/PAR-6/aPKC complex suppress the formation of epithelial junctions; this contrasts with the previous observation that the complex containing PAR-3 promotes it.
    Conclusions: These results indicate that PAR-6/aPKC selectively interacts with either mLgI or PAR-3 under the control of aPKC activity to regulate epithelial cell polarity.

    DOI: 10.1016/S0960-9822(03)00244-6

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  • Regulated protein-protein interaction between aPKC and PAR-3 plays an essential role in the polarization of epithelial cells 査読

    Y Nagai-Tamai, K Mizuno, T Hirose, A Suzuki, S Ohno

    GENES TO CELLS   7 ( 11 )   1161 - 1171   2002年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING LTD  

    Background: Recent studies have revealed that aPKC (atypical protein kinase C), PAR-3 and PAR-6 play indispensable roles in the regulation of various cell polarization events, from worms to mammals, suggesting that they comprise an evolutionarily conserved protein machinery which is essential for cell polarization. The three proteins interact with each other to form a ternary complex and thus mutually regulate their functionality and localization. Here, we investigated the biochemical nature of the aPKC-PAR-3 interaction in detail to clarify its functional importance in cell polarity.
    Results: The highly conserved 26 amino acid sequence 816-841, in PAR-3 was found to be necessary and sufficient for the tight association with aPKC. Among several conserved serine/threonine residues within the region, aPKC preferentially phosphorylates serine-827 in vitro , and this phosphorylation reduces the stability of the PAR-3-aPKC interaction. Several analyses using a phospho-serine 827 specific antibody have established that this phosphorylation by aPKC occurs in vivo . Over-expression of a point mutant of PAR-3 (S827A), which is predicted to form a stable complex with aPKC, causes defects in the cell-cell contact-induced cell polarization of epithelial MDCK cells, similarly to a dominant negative mutant of aPKC.
    Conclusion: These results imply that serine 827 in the aPKC binding site of PAR-3 is a target of aPKC and that the regulated interaction between a protein kinase, aPKC, and its substrate, PAR-3, plays an essential role in the establishment of cell polarity.

    DOI: 10.1046/j.1365-2443.2002.00590.x

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  • Over-expression of PAR-3 suppresses contact-mediated inhibition of cell migration in MDCK cells 査読

    A Mishima, A Suzuki, M Enaka, T Hirose, K Mizuno, T Ohnishi, H Mohri, Y Ishigatsubo, S Ohno

    GENES TO CELLS   7 ( 6 )   581 - 596   2002年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING LTD  

    Background: PAR-3 is one of the PAR proteins, previously named ASIP which are indispensable for the establishment of cell polarity in the embryo as well as differentiated epithelial cells. In mammalian epithelial cells, it forms a ternary complex with aPKC and PAR-6, and is localized to the tight junction that has been suggested as being important for creating cell polarity.
    Results: To gain insights into the mode of PAR-3 function in mammalian epithelial cells, we examined the effect of PAR-3 over-expression in MDCK cells. Although exogenous PAR-3-expression does not affect the epithelial polarity of confluent cells, it drastically transforms the morphology of cells at low density into a fibroblastic form with developed membrane protrusions. Time-lapse observations have revealed that PAR-3 over-expressing cells show intense motility, even after they have assembled into loose colonies, suggesting that the contact-mediated inhibition of cell migration (CIM) is suppressed. The expressions of E-cadherin and vimentin do not change with PAR-3 over-expression, suggesting that exogenous PAR-3 only disturbs the endogenous equilibrium of cellular states between a fundamental fibroblastic structure and an epithelial one. The co-expression of a dominant negative mutant of Rac1 and the addition of nocodazole strongly antagonize the effect of PAR-3 over-expression, suggesting the involvement of Rac1 activation and microtubule polymerizations.
    Conclusions: The data presented here suggest an intriguing link between the contact-mediated inhibition of cell migration and the regulation of cell polarity. The putative PAR-3 activities demonstrated here may function endogenously in the epithelial cell polarization process by being sequestered from the cytosol to the cell-cell junctional regions with aPKC and PAR-6 upon cell-cell adhesion.

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  • Dynamic changes in protein components of the tight junction during liver regeneration 査読

    Y Takaki, S Hirai, N Manabe, Y Izumi, T Hirose, M Nakaya, A Suzuki, K Mizuno, K Akimoto, S Tsukita, T Shuin, S Ohno

    CELL AND TISSUE RESEARCH   305 ( 3 )   399 - 409   2001年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER-VERLAG  

    The construction of the hepatocyte tight junction is one of the most important events during liver regeneration leading to the reorganization of the bile canaliculi and the repolarization of hepatocytes after cell division. To understand this event at the molecular level, we examined the expression of tight junction proteins by Western blot analysis and their cellular localization by immunofluorescence microscopy in regenerating rat liver after two-thirds hepatectomy. The levels of tight junction components such as claudin-3, ZO-1 and atypical protein kinase C (PKC)-specific interacting protein (ASIP) increased two- to three-fold over control levels in coordination with a peak 2-3 days after partial hepatectomy, whereas occludin levels remained unchanged. The bile canaliculi outlined by tight junction components and actin filaments reveal significant morphological changes from 2-3 days after partial hepatectomy. During this period, claudin-3/ZO-1 and ASIP/ZO-1 were nearly co-localized, whereas occludin was locally reduced or almost absent on the bile canaliculi outlined by ZO-1 staining. The uncoupled localization of F-actin and tight junction components was often observed. The function of hepatocytes, as revealed by the serum bile acids level, was distorted temporally at an early stage of regeneration but mostly restored 3 days after partial hepatectomy. These observations suggest that the de novo construction of tight junctions proceeds mainly 2-3 days after partial hepatectomy in parallel with the cell polarization required for hepatocyte function. However, the complete normalization of the composition of the tight junction components, such as occludin and the association with F-actin, requires additional time, which may support the regeneration of fully polarized normal hepatocytes.

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  • PAR-6 regulates aPKC activity in a novel way and mediates cell-cell contact-induced formation of the epithelial junctional complex 査読

    T Yamanaka, Y Horikoshi, A Suzuki, Y Sugiyama, K Kitamura, R Maniwa, Y Nagai, A Yamashita, T Hirose, H Ishikawa, S Ohno

    GENES TO CELLS   6 ( 8 )   721 - 731   2001年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL SCIENCE LTD  

    Background: PAR-6, aPKC and PAR-3 are polarity proteins that co-operate in the establishment of cell polarity in Caenorhabditis elegans and Drosophila embryos. We have recently shown that mammalian aPKC is required for the formation of the epithelia-specific cell-cell junctional structure. We have also revealed that a mammalian PAR-6 forms a ternary complex with aPKC and ASIP/PAR-3, and localizes at the most apical end of the junctional complex in epithelial cells.
    Results: The ternary complex formation and junctional co-localization of PAR-6 with aPKC and ASIP/PAR-3 are observed during the early stage of epithelial cell polarization. In addition, over-expression of the PAR-6 mutant with CRIB/PDZ domain in MDCK cells disturbs the cell-cell contact-induced junctional localization of tight junction proteins, as well as inhibiting TER development. Furthermore, the binding of Cdc42:GTP to the CRIB/PDZ domain of PAR-6 enhances the kinase activity of PAR-6-bound aPKC. Detailed analyses suggest that the binding of PAR-6 to aPKC has the intrinsic potential to activate aPKC, which is only released when Cdc42:GTP binds to the CRIB/PDZ domain.
    Conclusion: The results indicate the involvement of PAR-6 in the aPKC function which is required for the cell-cell adhesion-induced formation of epithelial junctional structures, possibly through the cooperative regulation of aPKC activity with Cdc42.

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  • The cell polarity protein ASIP/PAR-3 directly associates with junctional adhesion molecule (JAM) 査読

    K Ebnet, A Suzuki, Y Horikoshi, T Hirose, MK Meyer zu Brickwedde, S Ohno, D Vestweber

    EMBO JOURNAL   20 ( 14 )   3738 - 3748   2001年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    The establishment and maintenance of cellular polarity are critical for the development of multicellular organisms. PAR (partitioning-defective) proteins were identified in Caenorhabditis elegans as determinants of asymmetric cell division and polarized cell growth. Recently, vertebrate orthologues of two of these proteins, ASIP/PAR-3 and PAR-6, were found to form a signalling complex with the small GTPases Cdc42/Rac1 and with atypical protein kinase C (PKC). Here we show that ASIP/PAR-3 associates with the tight-junction-associated protein junctional adhesion molecule (JAM) in vitro and in vivo. No binding was observed with claudin-1, -4 or -5. In fibroblasts and CHO cells overexpressing JAM, endogenous ASIP is recruited to JAM at sites of cell-cell contact. Overexpression of truncated JAM lacking the extracellular part disrupts ASIP/PAR-3 localization at intercellular junctions and delays ASIP/PAR-3 recruitment to newly formed cell junctions. During junction formation, JAM appears early in primordial forms of junctions. Our data suggest that the ASIP/PAR-3-aPKC complex is tethered to tight junctions via its association with JAM, indicating a potential role for JAM in the generation of cell polarity in epithelial cells.

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  • Atypical protein kinase C is involved in the evolutionarily conserved PAR protein complex and plays a critical role in establishing epithelia-specific junctional structures 査読

    A Suzuki, T Yamanaka, T Hirose, N Manabe, K Mizuno, M Shimizu, K Akimoto, Y Izumi, T Ohnishi, S Ohno

    JOURNAL OF CELL BIOLOGY   152 ( 6 )   1183 - 1196   2001年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ROCKEFELLER UNIV PRESS  

    We have previously shown that during early Caenorhabditis elegans embryogenesis PKC-3, a C. elegans atypical PKC (aPKC), plays critical roles in the establishment of cell polarity required for subsequent asymmetric cleavage by interacting with PAR-3 [Tabuse, Y., Y. Izumi, F. Piano, K.J. Kemphues, J. Miwa, and S. Ohno. 1998. Development (Camb.). 125:3607-3614]. Together with the fact that aPKC and a mammalian PAR-3 homologue, aPKC-specific interacting protein (ASIP), colocalize at the tight junctions of polarized epithelial cells (Izumi, Y., H. Hirose, Y. Tamai, S.-I. Hirai, Y. Nagashima, T. Fujimoto, Y. Tabuse, K.J. Kemphues, and S, Ohno. 1998. J. Cell Biol. 143:95-106), this suggests a ubiquitous role for aPKC in establishing cell polarity in multicellular organisms. Here. we show that the overexpression of a dominant-negative mutant of aPKC (aPKCkn) in MDCK II cells causes mislocalization of ASIP/PAR-3. Immunocytochemical analyses, as well as measurements of paracellular diffusion of ions or non-ionic solutes, demonstrate that the biogenesis of the tight junction structure itself is severely affected in aPKCkn-expressing cells. Furthermore, these cells show increased interdomain diffusion of fluorescent lipid and disruption of the polarized distribution of Na+, K+-ATPase, suggesting that epithelial cell surface polarity is severely impaired in these cells. On the other hand, we also found that aPKC associates not only with ASIP/PAR-3, but also with a mammalian homologue of C. elegans PAR-6 (mPAR-6). and thereby mediates the formation of an aPKC-ASIP/PAR-3-PAR-6 ternary complex that localizes to the apical junctional region of MDCK cells. These results indicate that aPKC is involved in the evolutionarily conserved PAR protein complex, and plays critical roles in the development of the junctional structures and apico-basal polarization of mammalian epithelial cells.

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  • Muscle develops a specific form of small heat shock protein complex composed of MKBP/HSPB2 and HSPB3 during myogenic differentiation 査読

    Y Sugiyama, A Suzuki, M Kishikawa, R Akutsu, T Hirose, MMY Waye, SKW Tsui, S Yoshida, S Ohno

    JOURNAL OF BIOLOGICAL CHEMISTRY   275 ( 2 )   1095 - 1104   2000年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Previously, we identified a new mammalian sHSP, MKBP, as a myotonic dystrophy protein kinase-binding protein, and suggested its important role in muscle maintenance (Suzuki, A., Sugiyama, Y., Hayashi, Y., Nyu-i, N., Yoshida, M., Nonaka, I., Ishiura, S., Arahata, K., and Ohno, S. (1998) J. Cell Biol. 140, 1113-1124). In this paper, we develop the former work by performing extensive characterization of five of the six sHSPs so far identified, that is, HSP27, alpha B-crystallin, p20, MKBP/HSPB2, and HSPB3, omitting lens-specific alpha A-crystallin. Tissue distribution analysis revealed that although each sHSP shows differential constitutive expression in restricted tissues, tissues that express all five sHSPs are only muscle-related tissues. Especially, the expressions of HSPB3, identified for the first time as a 17-kDa protein in this paper, and MKBP/HSPB2 are distinctly specific to muscles. Moreover, these sHSPs form an oligomeric complex with an apparent molecular mass of 150 kDa that is completely independent of the oligomers formed by HSP27, alpha B-crystallin, and p20. The expressions of MKBP/HSPB2 and HSPB3 are induced during muscle differentiation under the control of MyoD, suggesting that the sHSP oligomer comprising MKBP/HSPB2 and HSPB3 represents an additional system closely related to muscle function. The functional divergence among sHSPs in different oligomers is also demonstrated in several ways: 1) an interaction with myotonic dystrophy protein kinase, which has been suggested to be important for the maintenance of myofibril integrity, was observed only for MKBP/HSPB2; 2) a myotube-specific association with actin bundles was observed for HSP27 and alpha B-crystallin, but not for MKBP/HSPB2; and 3) sHSPs whose mRNAs are induced by heat shock are alpha B-crystallin and HSP27. Taken together, the results suggest that muscle cells develop two kinds of stress response systems composed of diverged sHSP members, and that these systems work independently in muscle maintenance and differentiation.

    DOI: 10.1074/jbc.275.2.1095

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  • An atypical PKC directly associates and colocalizes at the epithelial tight junction with ASIP, a mammalian homologue of Caenorhabditis elegans polarity protein PAR-3 査読

    Y Izumi, T Hirose, Y Tamai, S Hirai, Y Nagashima, T Fujimoto, Y Tabuse, KJ Kemphues, S Ohno

    JOURNAL OF CELL BIOLOGY   143 ( 1 )   95 - 106   1998年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ROCKEFELLER UNIV PRESS  

    Cell polarity is fundamental to differentiation and function of most cells. Studies in mammalian epithelial cells have revealed that the establishment and maintenance of cell polarity depends upon cell adhesion, signaling networks, the cytoskeleton, and protein transport. Atypical protein kinase C (PKC) isotypes PKC zeta and PKC lambda have been implicated in signaling through lipid metabolites including phosphatidylinositol 3-phosphates, but their physiological role remains elusive. In the present study we report the identification of a protein, ASIP (atypical PKC isotype-specific interacting protein), that binds to aPKCs, and show that it colocalizes with PKC lambda to the cell junctional complex in cultured epithelial MD CKII cells and rat intestinal epithelia. In addition, immunoelectron microscopy revealed that ASIP localizes to tight junctions in intestinal epithelial cells. Furthermore, ASIP shows significant sequence similarity to Caenorhabditis elegans PAR-3. PAR-3 protein is localized to the anterior periphery of the one-cell embryo, and is required for the establishment of cell polarity in early embryos. ASIP and PAR-3 share three PDZ domains, and can both bind to aPKCs. Taken together, our results suggest a role for a protein complex containing ASIP and aPKC in the establishment and/or maintenance of epithelial cell polarity. The evolutionary conservation of the protein complex and its asymmetric distribution in polarized cells from worm embryo to mammalian-differentiated cells may mean that the complex functions generally in the organization of cellular asymmetry.

    DOI: 10.1083/jcb.143.1.95

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MISC

  • メディエーター複合体の液滴による転写ユニティー制御機構の解明

    鈴木秀文, 阿部竜太, 池陽子, 古郡華月, 小川真太郎, 豊田敦, 鈴木穣, 井野洋子, 木村弥生, 秋山智彦, 石川博子, 廣瀬智威, 山本達郎, 斉藤典子, 山口雄輝, 高橋秀尚

    日本生化学会大会(Web)   96th   2023年

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  • Differential roles of atypical protein kinase C isoforms in wound healing

    S. Osada, N. Noguchi, T. Hirose, T. Suzuki, M. Kagaya, K. Chida, S. Ohno, M. Manabe

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   137 ( 10 )   S306 - S306   2017年10月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE INC  

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  • 神経上皮細胞の一次繊毛におけるヘッジホッグ・シグナル伝達系の調節機構に対するPAR3の役割

    廣瀬智威, 廣瀬智威, 風間宏美, 野田哲生, 大野茂男

    日本生化学会大会(Web)   90th   ROMBUNNO.3LBA‐071 (WEB ONLY) - 071]   2017年

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    記述言語:日本語   出版者・発行元:生命科学系学会合同年次大会運営事務局  

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  • 筋サテライト細胞における細胞極性因子PAR3およびPKCiotaの役割

    瀬古大暉, 北嶋康雄, 廣瀬智威, 秋本憲和, 大野茂男, 小野悠介

    日本生化学会大会(Web)   90th   ROMBUNNO.2LBA‐089 (WEB ONLY)   2017年

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  • 一次繊毛におけるヘッジホッグ・シグナル伝達系の調節機構に対する細胞極性制御因子の関係

    廣瀬智威, 風間宏美, 藤田龍平, 大野茂男

    日本生化学会大会(Web)   88th   3P0193 (WEB ONLY) - [3P0193]   2015年

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    記述言語:日本語   出版者・発行元:(公社)日本生化学会  

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  • 足細胞の生化学

    佐藤 大輔, 廣瀬 智威, 大野 茂男

    生体の科学   63 ( 3 )   243 - 250   2012年5月

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    記述言語:日本語   出版者・発行元:金原一郎記念医学医療振興財団  

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    その他リンク: http://search.jamas.or.jp/link/ui/2012259093

  • Function of the polarity protein Par3 in Ras-induced skin tumorigenesis

    Sandra Iden, Helma van Riel, Ji-Ying Song, Tomonori Hirose, Shigeo Ohno, John G. Collard

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   131   S21 - S21   2011年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:NATURE PUBLISHING GROUP  

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  • スリット膜のターンオーバー制御を担う分子機構の解明 aPKC-Par複合体を介したnephrinの細胞膜局在制御

    佐藤 大輔, 張田 豊, 大門 主税, 栗原 秀剛, 廣瀬 智威, 大野 茂男

    日本腎臓学会誌   53 ( 3 )   377 - 377   2011年5月

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    記述言語:日本語   出版者・発行元:(一社)日本腎臓学会  

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  • ヒトアンジオテンシン1型受容体結合因子の腎内発現とIgA腎症がその発現に与える影響(Intrarenal Expression of Human Ang II Type 1 Receptor Interacting Molecule and Effects of IgA Nephropathy on Its Expression)

    増田 真一朗, 田村 功一, 涌井 広道, 前田 晃延, 出島 徹, 廣瀬 智威, 豊田 雅夫, 鈴木 大輔, 東 公一, 大澤 正人, 金岡 知彦, 柳 麻衣, 三橋 洋, 白 善雅, 小豆島 健護, 戸谷 義幸, 常田 康夫, 梅村 敏

    日本腎臓学会誌   53 ( 3 )   392 - 392   2011年5月

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    記述言語:英語   出版者・発行元:(一社)日本腎臓学会  

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  • Expression of angiotensin II type 1 receptor-interacting molecule in normal human kidney and IgA nephropathy

    Shin-Ichiro Masuda, Kouichi Tamura, Hiromichi Wakui, Akinobu Maeda, Toru Dejima, Tomonori Hirose, Masao Toyoda, Koichi Azuma, Masato Ohsawa, Tomohiko Kanaoka, Mai Yanagi, Shin-Ichiro Yoshida, Hiroshi Mitsuhashi, Miyuki Matsuda, Tomoaki Ishigami, Yoshiyuki Toya, Daisuke Suzuki, Yoji Nagashima, Satoshi Umemura

    American Journal of Physiology - Renal Physiology   299 ( 4 )   F720 - F731   2010年10月

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    記述言語:英語  

    The intrarenal renin-angiotensin system plays a crucial role in the regulation of renal circulation and sodium reabsorption through the activation of vascular, glomerular, and tubular angiotensin II type 1 (AT1) receptor signaling. We previously cloned a molecule that specifically interacted with the murine AT1 receptor to inhibit AT1 receptor signaling, which we named ATRAP (for AT1 receptor-associated protein). Since murine ATRAP was shown to be highly expressed in the kidney, in the present study we investigated expression and distribution of human ATRAP in normal kidney and renal biopsy specimens from patients with IgA nephropathy. In the normal human kidney, both ATRAP mRNA and protein were widely and abundantly distributed along the renal tubules from Bowman's capsule to the medullary collecting ducts. In all renal tubular epithelial cells, the ATRAP protein colocalized with the AT1 receptor. In renal biopsy specimens with IgA nephropathy, a significant positive correlation between ATRAP and AT 1 receptor gene expression was observed. There was also a positive relationship between tubulointerstitial ATRAP expression and the estimated glomerular filtration rate in patients with IgA nephropathy. Furthermore, we examined the function of the tubular AT1 receptor using an immortalized cell line of mouse distal convoluted tubule cells (mDCT) and found that overexpression of ATRAP by adenoviral gene transfer suppressed the angiotensin II-mediated increases in transforming growth factor-β production in mDCT cells. These findings suggest that ATRAP might play a role in balancing the renal renin-angiotensin system synergistically with the AT 1 receptor by counterregulatory effects in IgA nephropathy and propose an antagonistic effect of tubular ATRAP on AT1 receptor signaling. Copyright © 2010 the American Physiological Society.

    DOI: 10.1152/ajprenal.00667.2009

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  • p53のアポトーシス誘発タンパク質(ASPP2)は上皮細胞極性を調節するPAR複合体に関与する(Apoptosis-Stimulating Protein of p53 (ASPP2) is involved in the PAR Complex to Regulate Epithelial Cell Polarity)

    叢 偉立, 廣瀬 智威, 山下 暁朗, 水野 恵子, 張田 豊, 平野 久, 大野 茂男

    日本細胞生物学会大会講演要旨集   62回   141 - 141   2010年5月

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    記述言語:英語   出版者・発行元:(一社)日本細胞生物学会  

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  • Function of the polarity protein Par3 in Ras-induced skin tumorigenesis

    S. Iden, H. van Riel, A. E. Mertens, J. -Y. Song, T. Hirose, S. Ohno, J. G. Collard

    EUROPEAN JOURNAL OF CELL BIOLOGY   89   4 - 4   2010年3月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER GMBH, URBAN & FISCHER VERLAG  

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  • Chronic angiotensin II infusion suppresses renal expression of angiotensin II type 1 receptor-interacting molecule ATRAP to decrease angiotensin II type 1 receptor internalization

    Kouichi Tamura, Hiromichi Wakui, Miyuki Matsuda, Toru Dejima, Atsu-ichiro Shigenaga, Shin-ichiro Masuda, Koichi Azuma, Akinobu Maeda, Tomohiko Kanaoka, Masato Ohsawa, Tomonori Hirose, Tomoaki Ishigami, Yoshiyuki Toya, Machiko Yabana, Satoshi Umemura

    ENDOCRINE JOURNAL   57   S558 - S558   2010年3月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:JAPAN ENDOCRINE SOC  

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  • 細胞極性遺伝子:マウスモデルから疾患の理解と診断・治療・予防へ

    大野茂男, 廣瀬智威, 秋本和憲, 山下暁朗, 鈴木厚, 平井秀一

    横浜医学   60 ( 1/2 )   84 - 90   2009年5月

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    記述言語:日本語  

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  • ポドサイトの細胞極性異常に基づく新しい巣状糸球体硬化症モデルマウス

    廣瀬智威, 佐藤大輔, 栗原秀剛, 日下智保, 廣瀬博子, 秋本和憲, 松阪泰二, 市川家國, 野田哲生, 大野茂男

    横浜医学   60 ( 1/2 )   (JA)161,(EN)161-162   2009年5月

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  • Genes relevant to cellular polarity: from mouse models to human diseases (第2回横浜市立大学・米国食品医薬品庁共催--国際学術ワークショップ報告 生物製剤の開発と新しい治療法のためのバイオマーカー)

    大野 茂男, 廣瀬 智威, 秋本 和憲

    横浜医学   60 ( 1 )   84 - 90   2009年5月

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    記述言語:英語   出版者・発行元:横浜市立大学医学会  

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  • 腎糸球体ポドサイトのスリット膜維持の新しい分子機構:細胞極性制御系aPKC‐PAR系の役割

    廣瀬智威, 佐藤大輔, 栗原秀剛, 日下智保, 廣瀬博子, 秋本和憲, 松阪泰二, 市川家國, 伊藤秀一, 野田哲生, 大野茂男

    日本腎臓学会誌   51 ( 3 )   264   2009年4月

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  • 細胞極性制御因子aPKCは腎糸球体ポドサイトにおけるスリット膜の機能に必須の役割を担う

    廣瀬智威, 佐藤大輔, 栗原秀剛, 松阪泰二, 秋本和憲, 市川家國, 野田哲生, 大野茂男

    生化学   2P-0651   2007年

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  • aPKC-PARシステム--普遍的な細胞極性制御機構 (〔2001年〕12月第5土曜特集 発生学から再生医療へ) -- (発生学の進歩 細胞極性)

    廣瀬 智威, 大野 茂男

    医学のあゆみ   199 ( 13 )   861 - 865   2001年12月

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    記述言語:日本語   出版者・発行元:医歯薬出版  

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    その他リンク: http://search.jamas.or.jp/link/ui/2002173748

  • aPKC λ conditional gene targeting miceの作製と機能解析

    秋本和憲, 中谷雅明, 杉谷善信, 広瀬智威, 山中ひとみ, 美野輪治, 唐沢美香, 大野茂男, 野田哲生

    日本分子生物学会年会プログラム・講演要旨集   23rd   680   2000年11月

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  • 低分子量熱ショックタンパク質(sHSP)群で構成される筋特異的なストレス耐性系

    杉山 由樹, 鈴木 厚, 岸川 優, 阿久津 里佳, 廣瀬 智威, 加藤 兼房, WAYE M., 大野 茂男

    日本分子生物学会年会プログラム・講演要旨集   21   578 - 578   1998年12月

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    記述言語:日本語  

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  • Atypical PKC特異的結合蛋白質の上皮組織における分布と細胞極性との関係

    廣瀬 智威, 泉 裕士, 長嶋 洋治, 江原 美智子, 藤本 豊士, 大野 茂男

    日本分子生物学会年会プログラム・講演要旨集   21   551 - 551   1998年12月

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    記述言語:日本語  

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  • 上皮細胞の細胞接着構造形成過程におけるASIP-aPKC複合体の果たす役割

    鈴木 厚, 大西 哲生, 泉 裕士, 廣瀬 智威, 野田 久美, 大野 茂男

    日本分子生物学会年会プログラム・講演要旨集   21   551 - 551   1998年12月

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  • ASIP(aPKC-specific interacting protein)を足場とするシグナル伝達複合体

    鈴木 友香里, 廣瀬 智威, 泉 裕士, 大野 茂男

    日本分子生物学会年会プログラム・講演要旨集   21   550 - 550   1998年12月

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  • aPKC群結合タンパク質の単離およびその生化学的解析

    田中純平, 秋本和憲, 山中智行, 中谷雅明, 吉田道彦, 広瀬智威, 鈴木厚, 田沼靖一, 大野茂男

    日本分子生物学会年会プログラム・講演要旨集   21st   550   1998年11月

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講演・口頭発表等

  • 教育講演:様々な細胞の極性を制御する共通分子機構 招待

    廣瀬智威

    第85回 日本皮膚科学会東京支部学術大会  2021年11月 

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    会議種別:口頭発表(招待・特別)  

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  • 大脳発生過程においてPAR3はヘッジホッグ・シグナリングの制御を介して神経前駆細胞の増殖に制限をかける

    廣瀬智威

    第45回日本分子生物学会年会、ポスター発表・サイエンスピッチ  2022年11月 

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  • 精子形成における転写伸長制御因子の機能解析

    劉凱文, 廣瀬智威, 髙橋秀尚

    第46回日本分子生物学会年会、ポスター発表・サイエンスピッチ  2023年12月 

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受賞

  • 医学研究奨励賞

    2011年4月   横浜市立大学医学会  

    廣瀬智威

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共同研究・競争的資金等の研究課題

  • 精原幹細胞の同期的な分化制御における統合的かつ同調的な遺伝子発現調節機構の解明

    研究課題/領域番号:23K06381  2023年4月 - 2026年3月

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    廣瀬 智威

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    配分額:4550000円 ( 直接経費:3500000円 、 間接経費:1050000円 )

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  • 大脳皮質形成における霊長類型の神経幹細胞産生に関わる遺伝子の網羅的探索

    研究課題/領域番号:20K06893  2020年4月 - 2023年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    廣瀬 智威

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    配分額:4420000円 ( 直接経費:3400000円 、 間接経費:1020000円 )

    霊長類型の神経幹細胞産生に関わる遺伝子を探索するため、これまでに終脳特異的PAR3欠損マウス(Par3 cKOマウス)で認められる同型の神経幹細胞の過剰産生に伴って発現変動する遺伝子をいくつか同定してきた。それらの遺伝子が関与するシグナル伝達系を制御する場として、一次シリアが中心的な役割を担うことが知られていたので、先ずこれに注目することにした。2020年度は、Par3 cKOマウスの神経幹・前駆細胞の一次シリアにおけるこの制御系の異常を捉えるべく、超解像顕微鏡を用いて当該のシグナル伝達タンパク質の定量的な局在解析を実施した。更に、当該マウスで認められる霊長類型神経幹細胞の過剰産生に伴って発現変動する候補遺伝子の網羅的探索を実施するため、十分な数の適切なマウス胚の準備を進めた。今後は、特に一次シリアの形成・機能・シグナル伝達系に関連する遺伝子に注目し、上記の超解像顕微鏡解析も応用しつつ、異常の評価・検証を行う。
    本研究の成果として、区切りとなる研究結果が得られたことから、論文投稿を前倒しで開始した。残念ながら2020年度中の採択は叶っていないものの、アピールポイントを明確化するなどの推敲を重ねており、採択を目指して注力する。
    並行して実施しているいくつかの共同研究でも進展が得られた。特に、ヒト大脳皮質形成異常の原因遺伝子の機能解析で参加した共同研究は、論文発表に結び付いた。更に別の共同研究(米国2件)に用いるため、自身で樹立したPAR3コンディショナルノックアウトマウス系統の空輸を完了した。

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  • 酸化ストレス応答による細胞極性の異常を起点とした病態発生の分子機構の解析

    研究課題/領域番号:16K08739  2016年4月 - 2019年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    堀越 洋輔, 大野 茂男, 廣瀬 智威

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    炎症性腸疾患では、炎症反応による酸化ストレスを伴い極性化した上皮細胞から成る管腔組織構造の破綻が確認される。しかし、酸化ストレスの細胞極性に及ぼす作用の詳細は不明であった。本研究では、「炎症性腸疾患で起こる管腔構造の破綻が、炎症反応を介した酸化ストレスによる aPKC の過剰な活性化を経た aPKC-PAR 複合体の形成阻害によって誘導される事を証明する」を目的とした。本研究課題により、炎症応答による酸化ストレスは、脂溶性情報伝達物であるPS(フォスファチジルセリン)の過酸化体を発生させ、aPKCの過剰活性化を引き起こし上皮組織の形態異常(極性異常)を引き起こす可能性が示唆された。

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  • 細胞極性制御因子aPKCによる毛包幹細胞の休眠制御機構

    研究課題/領域番号:15K09755  2015年4月 - 2018年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    長田 真一, 能登 舞, 廣瀬 智威, 鈴木 倫子, 加賀谷 昌美

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    表皮には細胞極性に関わる2種類のaPKC (atypical protein kinase C) 分子種(aPKCζ, aPKCλ)が発現している。本研究で私たちは、創傷治癒と創傷後毛包新生という、皮膚恒常性の維持に関わる重要な過程に、これらのaPKC分子種がどのように関与しているのかを調べた。表皮特異的aPKCλ欠損マウスでは創傷治癒が遅延し、Wnt経路の活性化を伴う創傷後毛包新生が亢進したが、aPKCζ欠損マウスではこのような現象は観察されなかった。本研究により、aPKCλが創傷治癒と創傷後毛包新生を繋ぐ重要な分子であることが初めて明らかになった。

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  • 組織幹前駆細胞の極性制御と運命決定

    研究課題/領域番号:23112003  2011年4月 - 2016年3月

    日本学術振興会  科学研究費助成事業 新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    大野 茂男, 廣瀬 智威, 佐々木 和教, 中谷 雅明, 飯田 直子, 佐藤 大輔, 佐藤 由典, 田村 可奈, 峰松 尚子, 髙栁 亜由美, 藤田 龍平

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    配分額:154050000円 ( 直接経費:118500000円 、 間接経費:35550000円 )

    本研究では、乳腺組織をモデルとして、細胞極性タンパク質(様々な細胞の極性を制御する普遍機構(aPKC-PAR系)が、乳腺組織幹細胞及び前駆細胞の自己亢進・増殖などにおける役割を解析しました。そして、幹細胞の自己更新にaPKCが正の役割を持っていること、さらにを媒介する新たな転写因子の同定に成功しました。一方、管腔前駆細胞では、aPKCはその抑制的な役割を果たしており、ErbB2の転写を抑制している事を見いだしました。それらの異なったタイプの乳がんで、上述の各々のケースがある事を示唆しました。診断に役立つことが期待されます。

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  • 細胞極性のシグナリングの分子・細胞機構

    研究課題/領域番号:22247030  2010年 - 2012年

    日本学術振興会  科学研究費助成事業 基盤研究(A)  基盤研究(A)

    大野 茂男, 平井 秀一, 鈴木 厚, 秋本 和憲, 山下 暁朗, 廣瀬 智威, 中谷 雅明, 佐々木 和教

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    配分額:44460000円 ( 直接経費:34200000円 、 間接経費:10260000円 )

    上皮細胞を初めとする様々な細胞の極性制御の要として働いている普遍的な細胞極性シグナル経路、PAR-aPKC 系の新規構成員として ASPP2 を発見し、細胞極性の制御と細胞死の制御の間の関係を示唆した。PAR-aPKC 系の新たな制御機構として、aPKC 結合タンパク質 KIBRA が aPKCのキナーゼ活性を基質と競合的に抑制し Lgl とは異なる機構で aPKC を通じたアピカル膜ドメイン形成のプロセスを特異的に抑制することを見いだした。

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  • 普遍的細胞極性制御装置による哺乳動物初期胚発生制御機構の研究

    研究課題/領域番号:21116004  2009年7月 - 2014年3月

    日本学術振興会  科学研究費助成事業 新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    鈴木 厚, 廣瀬 智威, 中谷 雅明

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    配分額:151970000円 ( 直接経費:116900000円 、 間接経費:35070000円 )

    哺乳動物の受精卵が身体作りをする非常に初期の段階(8から32細胞の時期)では、細胞塊の外側の細胞が次第に上皮細胞としての性質を獲得することが、細胞の分化状態がゆっくりと固定化していくために非常に重要である。本研究では、哺乳類初期発生にとって細胞の上皮化、脱上皮化が決定的に重要であることに着目し、近年明らかになってきた普遍的な極性制御因子群(PAR-aPKCタンパク質群)が細胞の上皮化を制御する機構を詳細に分析した。その結果、これまで知られていなかった新しい制御機構を3点にわたって見出すに至り、今後の哺乳類初期発生過程解明の基盤となる重要な知見を生み出した。

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担当経験のある科目(授業)

  • 医科学概論(分子生物学:分子生物学的手法を駆使した糸球体疾患の病態機構解明)

    機関名:横浜市立大学大学院医学研究科

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  • 医科学実験法(細胞株を用いた遺伝子組換え実験法、PCR実験法)

    機関名:横浜市立大学医学研究科

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  • 代謝障害・内分泌異常、体液と電解質異常・酸塩基平衡異常

    機関名:横浜市救急救命士養成科

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  • 医科学概論(細胞生物学:細胞増殖と細胞分化、細胞運動とその制御)

    機関名:横浜市立大学大学院医学研究科

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  • 分子細胞生物学(細胞の観察・分画・培養、細胞及び個体の遺伝子操作、細胞膜の構造、幹細胞の維持と分化、細胞の誕生・分化・死)

    機関名:横浜市立大学医学部医学科

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  • 看護生化学(生体構成成分と 細胞の構成;遺伝情報とその発現)

    機関名:横浜市立大学医学部看護学科

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  • 分子生物学論文演習(PBL/TBL形式での原著論文読解と発表会)

    機関名:横浜市立大学医学部医学科

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  • 分子生物学論文演習(2人/組の個別指導による原著論文読解と発表会)

    機関名:横浜市立大学医学部医学科

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  • 分子生物学実習 (1. PCR法を用いたゲノムDNAの解析とPoint Mutationの導入;2. 大腸菌を用いたGST融合蛋白質の発現・精製・検出)

    機関名:横浜市立大学医学部医学科

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