細胞接着の異常と発がん・転移機構

• Project/Area Number: 06283225
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: Kyoto University (1995-1999); Okazaki National Research Institutes (1994)
• Principal Investigator: 月田 承一郎 京都大学, 医学研究科, 教授 (50155347)
• Co-Investigator(Kenkyū-buntansha): 月田 早智子 京都大学, 医療技術短期大学部, 教授 (00188517) ; 米村 重信 京都大学, 医学研究科, 講師 (60192811) ; 永渕 昭良 京都大学, 医学研究科, 講師 (80218023) ; 伊藤 雅彦 京都大学, 医学研究科, 助手 (70270486)
• Project Period (FY): 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥215,000,000 (Direct Cost: ¥215,000,000); Fiscal Year 1999: ¥41,000,000 (Direct Cost: ¥41,000,000); Fiscal Year 1998: ¥40,000,000 (Direct Cost: ¥40,000,000); Fiscal Year 1997: ¥39,000,000 (Direct Cost: ¥39,000,000); Fiscal Year 1996: ¥35,000,000 (Direct Cost: ¥35,000,000); Fiscal Year 1995: ¥30,000,000 (Direct Cost: ¥30,000,000); Fiscal Year 1994: ¥30,000,000 (Direct Cost: ¥30,000,000)
• Keywords: オクルディン / クローディン / ノックアウトマウス / タイトジャンクション / アドヘレンスジャンクション / カテニン / ERM蛋白質 / カドヘリン / ZO-1 / エズリン / ラディキシン / モエシン / ノックアウト / がん / αカテニン / Rho / Rhoキナーゼ / ERM / 細胞接着 / 発がん / 転移 / βカテニン / ZO-2 / 細胞間接着 / 情報伝達機構 / 癌抑制遺伝子 / ES細胞 / ラディチシン

Research Abstract

第1プロジェクト
TJの接着分子として同定されたオクルディンとクローディン分子群についての解析を中心に行った。
(1)一昨年までにオクルディンのノックアウトマウスを作製することに成功しており、予備的な解析の結果、慢性の胃炎を引き起こし、前癌状態ともいえる状態にまで至ることが分かっていた。このマウスを徹底的に解析し、細胞癌化とTJの関係について調べた。
(2)クローディンは現在のところ16種類存在することが分かってきたが、それぞれの遺伝子を破壊したマウスの作製に着手した。
第2プロジェクト
AJとTJの新たな裏打ち蛋白質の同定と、これまでに同定された裏打ち蛋白質の機能解析を目指した。
(1)βカテニンの分解が細胞癌化にとって重要な役割を果たしていることは、最近広く認められるところであるが、αカテニンの分解については解析されていない。αカテニンの分解も精密に制御されていることが、予備的解析により明らかになっていたので、その分子機構を探った。
(2)ERM蛋白質の機能解析の一環としてノックアウトマウスの作製を行っていた。すでに、モエシンノックアウトマウスの解析は終了していた。現在、ラディキシンノックアウトマウスが生まれてきたところであり、このマウスの解析を進めている。
(3)TJの裏打ち蛋白質ZO-1、-2、-3の機能解析のためにノックアウトマウスの作製を開始した。

Research Products

• [Publications] Yonemura, S., Tsukita, Sa., and Tsukita, Sh.: "Direct involvement of ezrin/radixin /moesin (ERM) binding membrane proteins in the organization of microvilli in collaboration with activated ERM proteins"J. Cell Biol.. 145. 1497-1506 (1999)
• [Publications] Morita, K., Sasaki, H., Fujimoto, K., Furuse, M., and Tsukita Sh.: "Claudin 11/OSP based tight junctions in myelinated sheaths of oligodendrocytes and in Sertoli cells in testis"J. Cell Biol.. 145. 579-588 (1999)
• [Publications] Furuse, M., Sasaki, H., and Tsukita, Sh.: "Manner of interaction of heterogenous claudin species within and between tight junction strands"J. Cell Biol.. 147. 891-903 (1999)
• [Publications] Itoh, M., Furuse, M., Morita, K., Kubota, K., and Tsukita, Sh.: "Molecular basis for the recruitment of three MAGUKs, ZO-1, ZO-2 and ZO-3 to tight junctions : Direct binding of their first PDZ domains to the COOH-termini of claudins"J. Cell Biol.. 147. 1351-1363 (1999)
• [Publications] Sonoda, N., Furuse, M., Sasaki, H., Yonemura, S., Katahira, J., Horiguchi, Y., and Tsukita, Sh.: "Clostridium perfringens enterotoxin fragment removes specific claudins from tight junction strands : Evidence for direct involvement of claudins in tight junction barrier"J. Cell Biol.. 147. 195-204 (1999)
• [Publications] Morita, K., Sasaki, H., Furuse, M. and Tsukita, Sh.: "Endothelial claudin : claudin-5/TMVCF constitutes tight junction strands in endothelial cells"J. Cell Biol.. 147. 185-194 (1999)
• [Publications] Yonemura,S.et al.: "ERM proteins bind to a specific group of integral membrane proteins containing a positively-harget amino acid cluster in their juxta-membrane cytoplasmic domain." J.Cell Biol.140. 885-895 (1998)
• [Publications] Matsui,T.et al.: "Rho-kinase phosphorylates carboxy-terminal threonines of ERM proteins and regulates their head-to-tail association" J.Cell Biol.140. 647-657 (1998)
• [Publications] Saitou,M.et al.: "Occludin-deficient embryonic stem cells can diferentiate into polarized epithelial cells bearing tight junctions" J.Cell Biol.141. 397-408 (1998)
• [Publications] Furuse,M.et al.: "Claudin-1 and -2: Novel integral membrane proteins localizing at tight junctions with no sequence similarity to occludin." J.Cell Biol.141. 1539-1550 (1998)
• [Publications] Furuse,M.et al.: "A single gene product,claudin-1 or -2,reconstitutes tight junction strands and recruits occludin in fibroblasts" J.Cell Biol.143. 391-401 (1998)
• [Publications] Morita,K.et al.: "Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands" Proc.Natl.Acad.Sci.USA. (印刷中). (1999)
• [Publications] Sachiko TSUKITA: "ERM proteins : Head-to-tail regulation of actin/plasma membrane regulations." Trends in Biological Science. 22. 53-58 (1997)
• [Publications] Sachiko TSUKITA: "ERM (Ezrin/Radixin/Moesin) family : From cytoskeleton to signal transduction" Current Opinion in Cell Biology. 9. 70-75 (1997)
• [Publications] Takahisa KONDO: "ERM (Ezrin/Radixin/Moesin)-based molecular mechanism of microvillar breakdown at an early stage of apoptosis." Journal of Cell Biology. 139. 758-767 (1997)
• [Publications] Takeshi MATSUI: "Rho-kinase phosphorylates carboxy-terminal threonines of ERM proteins and regulates their head-to-tail association." Journal of Cell Biology. (1998)
• [Publications] Yoshinori DOI: "Moesin is not a receptor for measles virus entry into mouse embryonic stem cells." Journal of Virology. (1998)
• [Publications] Shigenobu Yonemura: "ERM proteins bind to a specific group of integral membrane proteins containing a positively-charged amino acid cluster in their juxta-membrane cytoplasmic domain." Journal of Cell Biology. (1998)
• [Publications] Furuse,M.et al.: "Overexpression of occludin,a tight junction-associated integral membrane protein,induces the formation of intracellular multilamellar bodies bearing tight junction-like structures" J.Cell Sci.109. 429-435 (1996)
• [Publications] Ando-Akatsuka,Y.et al.: "Interspecies diversity of the occludin sequence : cDNA cloning of human,mouse,dog and ratkangaroo homologues" J.Cell Biol.133. 43-47 (1996)
• [Publications] Tokai,N.et al.: "Kid,a novel kinesin-like DNA binding protein,is localized to chromosomes and the mitotic spindle" EMBO J.15. 457-467 (1996)
• [Publications] Hirase,T.et al.: "A 155kDa undercoat-constitutive protein of cell-to-cell adherens junctions" Eur.J.Cell Biol.72. 174-181 (1997)
• [Publications] Hirase,M.et al.: "Regulation Mecanism of ERM(Ezrin/Radixin/Moesin) protein/Plasma Membrane Association : Possible Involvement of Phosphatidylinositol Turnover and Rho-dependent Signaling Pathway" J.Cell Biol. 135. 1-16 (1996)
• [Publications] McCarthy.et al.: "Occludin is a functional component of the tight juncton." J.Cell Sci.109. 2287-2298 (1996)
• [Publications] Yonemura, S. et. al.: "Cell-to-cell adherens junction formation and actin filament organization: Similarities and difference between non-polarized fibroblasts and polarized epithelial cells." J. Cell Sci.108. 127-142 (1995)
• [Publications] Oyama, T. et. al.: "A truncated beta-catenin disrupts the interaction between E-cadherin and alpha-catenin : A cause of loss of intercellular adhesiveness in human cancer cell lines" Cancer Res.54. 6282-6287 (1995)
• [Publications] Takeda, H. et. al.: "V-src kinase shifts the cadherin-based cell adhesion from the strong to the weak state and beta-catenin is not required for the shift." J. Cell Biol.131. 1839-1847 (1995)
• [Publications] Furuse, M. et. al.: "Overexpression of occludin, a tight junction-associated integral membrane protein, induces the formation of intracellular multilamellar bodies bearing tight junction-like structures" J. Cell Sci.109. 429-425 (1996)
• [Publications] Tokai, N. et. al.: "Kid, a novel kinesin-like DNA binding protein, is localized to chromosomes and the mitotic spindle" EMBO J.15(in press). (1996)
• [Publications] Ando-Akatsuka, Y. et. al.: "Interspecies diversity of the occludin sequence:cDNA cloning of human, mouse, dog and rat-kangaroo homologues" J. Cell Biol.(in press). (1996)
• [Publications] Kasahara,H.: "Autoimmune myocarditis induced in mice by c-protein." J.Clin.Invest.94. 1026-1036 (1994)
• [Publications] Nagafuchi,A.: "The nervous tissue-specific loss of the expression of alpha-catenin,the 102kD cadherin-associated protein,during development." Develop,Growth & Differ.36. 59-71 (1994)
• [Publications] Takeuchi,K.: "Perturbation of cell adhesion and microvilli formation by antisense oligonuclotides to ERM family members." J.Cell Biol.125. 1371-1384 (1994)
• [Publications] Takeuchi,K.: "Structural diversity of band 4.1 superfamily members." J.Cell Sci.107. 1921-1928 (1994)
• [Publications] Tsukita,S.: "ERM family members as molecular linkers between the cell surface glycoprotein CD44 and actin-based cytoskeletons." J.Cell Biol.126. 391-401 (1994)
• [Publications] Nagafuchi,A.: "The roles of catenins in the cadherin-mediated cell adhesion:Functional analysis of E-cadherin-alpha catenin fusion molecules." J.Cell Biol.127. 602-613 (1994)