1997 Fiscal Year Annual Research Report

TGF-b-DPC-4による細胞周期制御機構の解析

Research Project

Project/Area Number	09470042
Research Institution	Osaka University
Principal Investigator	秋山徹大阪大学, 微生物病研究所, 教授 (70150745)
Co-Investigator(Kenkyū-buntansha)	石館宇夫東京大学分子細胞生物学研究所, 助手
Keywords	DPC-4, / TGF-b, / TAK1, / TAB-1, / 増殖制御、 / 細胞周期、 / シグナル伝達、 / 転写制御
Research Abstract	DPC-4を強制発現すると細胞増殖を強力に抑制することを見出したので細胞周期上での作用点を解析しG1期からS期への移行を阻害することを明らかにした。膵臓癌で見出された変異をもつDPC-4はこのような活性を示さなかった。一方、やはりTGF-bの下流でシグナル伝達に関与していると考えられるTAK1は強制発現しても顕著な細胞周期制御活性は示さなかった。TAK1とTAB1を同時に強制発現した場合やN端が欠失して活性化したTAK1を強制発現した場合にも顕著な効果はみられなかった。さらに、DPC-4による細胞周期制御機構を明らかにするためにyeastを用いたtwo-hybrid systemによりDPC-4と結合する細胞蛋白質を検索し、_sMAD2およびMCM7が複合体を形成する可能性のあることを見出した。SMDA2についてはin vitroでのpull-down assayおよびin vivoでの免疫沈降-Western blottingにより実際に細胞内でDPC-4と結合していることが明かとなった。一方MCM7についてははっきりした結果は得られず細胞内では結合していないのではないかと考えられた。DPC-4の作用の解明にはその標的遺伝子の解明が最も重要であると考えられるが一つの候補として_p21遺伝子が見出された。すなわちDPC-4はTAK1と相乗的にp21のプロモーターに作用して活性化することが見出された。さらにDPC-4をinducibleに発現する細胞株の作製を試み、DPC-4のより重要な標的遺伝子をsubtraction法およびdifferential display法により見出すための準備をおこなった。

Research Products
(16 results)

All Other

All Publications (16 results)

[Publications] K.Satoh et al.: "DAP-1,a novel proteinthat interacts with the guanylate kinase-like domains of DLG and PSD-95." Genes to Cells. 2. 415-424 (1997)
[Publications] T.Ishidate et al.: "Identification of a novel nuclear localization signal in Sam68." FEBS LETTERS. 409. 237-241 (1997)
[Publications] M.Moriyama et al.: "The BCL-6 protein is phosphorylated at multiple sites in its serine-and proline-clustered region by mitogen activated protein kinase(MAPK)in vivo." Oncogene. 14. 2465-2474 (1997)
[Publications] T,Kiyono et al.: "Binding of high risk HPV E6 oncoproteins to PDZ-domain-containing proteins." Proc. Natl. Acad. Sci. USA. 94. 11616-11612 (1997)
[Publications] Y.Murata et al.: "The Wilms tumor suppressor gene WT1 induces G1 arrest and apoptosis in myeloblastic leukemia M1 cells." FEBS LLETTERS.409. 41-45 (1997)
[Publications] H.Shibata et al.: "Rapid colorectal adenoma formation initiated by conditional targeting of the Apc gene." Science. 278. 120-123 (1997)
[Publications] T.Senda et al.: "Expression and localization of the APC tumor suppressor protein in the nervous system." Neuroscience. 83. 857-866 (1998)
[Publications] T.Yamagami et al.: "Suppression of Wilms tumor gene(WT1)expression induces G2/M arest in leukemic cells." Leukemia Res.(in press).
[Publications] T.Senda et al.: "Association of the MCC(Multated in colorectal cancer)gene product to the plasma membrane and membrane organelles." Medical Electron Microscopy. (in press).
[Publications] K.Inoue et al.: "Wilms tumor gene(WT1) competes with differentiation-induced signal in hematopoietic progenitor cells." Blood. (in press).
[Publications] M.Takemura et al.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase" Oncogene. 15. 2483-2492 (1997)
[Publications] Y.Dobashi et al.: "The Wilms tumor protein is persistently associated with the nuclear matrix throughout the cell cycle." Mol. Cell. Biochem.171. 121-126 (1997)
[Publications] I.Miyashiro et al.: "Midkine expression in human breast cancers: Expression of truncated form." Breast Can. Res. Treatment. 43. 1-6 (1997)
[Publications] K.Kagawa et al.: "Overexpression of GML promotes radiation-induced cell cycle arrest and apoptosis." Biochem. Biophys. Res. Commun.241. 481-485 (1997)
[Publications] Y.Dobashi et al.: "Cell cycle control with minimal participation of CDK2 in a murine fibrosarcoma clone cultured in protein-free medium." Biochem. Biophys. Res. Commun.232. 622-625 (1997)
[Publications] J.Adachi et al.: "Growth suppression of non-small cell lung carcinoma cells by the introduction of the p16INK4A gene." Int. J. Cancer. 10. 33-39 (1997)

1997 Fiscal Year Annual Research Report

TGF-b-DPC-4による細胞周期制御機構の解析

Principal Investigator

秋山 徹 大阪大学, 微生物病研究所, 教授 (70150745)

Research Products

[Publications] K.Satoh et al.: "DAP-1,a novel proteinthat interacts with the guanylate kinase-like domains of DLG and PSD-95." Genes to Cells. 2. 415-424 (1997)

[Publications] T.Ishidate et al.: "Identification of a novel nuclear localization signal in Sam68." FEBS LETTERS. 409. 237-241 (1997)

[Publications] M.Moriyama et al.: "The BCL-6 protein is phosphorylated at multiple sites in its serine-and proline-clustered region by mitogen activated protein kinase(MAPK)in vivo." Oncogene. 14. 2465-2474 (1997)

[Publications] T,Kiyono et al.: "Binding of high risk HPV E6 oncoproteins to PDZ-domain-containing proteins." Proc. Natl. Acad. Sci. USA. 94. 11616-11612 (1997)

[Publications] Y.Murata et al.: "The Wilms tumor suppressor gene WT1 induces G1 arrest and apoptosis in myeloblastic leukemia M1 cells." FEBS LLETTERS.409. 41-45 (1997)

[Publications] H.Shibata et al.: "Rapid colorectal adenoma formation initiated by conditional targeting of the Apc gene." Science. 278. 120-123 (1997)

[Publications] T.Senda et al.: "Expression and localization of the APC tumor suppressor protein in the nervous system." Neuroscience. 83. 857-866 (1998)

[Publications] T.Yamagami et al.: "Suppression of Wilms tumor gene(WT1)expression induces G2/M arest in leukemic cells." Leukemia Res.(in press).

[Publications] T.Senda et al.: "Association of the MCC(Multated in colorectal cancer)gene product to the plasma membrane and membrane organelles." Medical Electron Microscopy. (in press).

[Publications] K.Inoue et al.: "Wilms tumor gene(WT1) competes with differentiation-induced signal in hematopoietic progenitor cells." Blood. (in press).

[Publications] M.Takemura et al.: "Phosphorylated retinoblastoma protein stimulates DNA polymerase" Oncogene. 15. 2483-2492 (1997)

[Publications] Y.Dobashi et al.: "The Wilms tumor protein is persistently associated with the nuclear matrix throughout the cell cycle." Mol. Cell. Biochem.171. 121-126 (1997)

[Publications] I.Miyashiro et al.: "Midkine expression in human breast cancers: Expression of truncated form." Breast Can. Res. Treatment. 43. 1-6 (1997)

[Publications] K.Kagawa et al.: "Overexpression of GML promotes radiation-induced cell cycle arrest and apoptosis." Biochem. Biophys. Res. Commun.241. 481-485 (1997)

[Publications] Y.Dobashi et al.: "Cell cycle control with minimal participation of CDK2 in a murine fibrosarcoma clone cultured in protein-free medium." Biochem. Biophys. Res. Commun.232. 622-625 (1997)

[Publications] J.Adachi et al.: "Growth suppression of non-small cell lung carcinoma cells by the introduction of the p16INK4A gene." Int. J. Cancer. 10. 33-39 (1997)

秋山徹大阪大学, 微生物病研究所, 教授 (70150745)