1991 年度実績報告書

細胞癌化における蛋白質リン酸化

研究課題

研究課題/領域番号	03151038
研究機関	大阪大学
研究代表者	秋山徹微生物病研究所, 発癌遺伝子部門, 助教授 (70150745)
研究分担者	西田栄介東京大学, 理学部, 助手 (60143369) 月田早智子岡崎国立共同研究機構, 生理学研究所・生体情報研究系液性情報 (00188517) 岡田雅人大阪大学蛋白質研究所, 代謝部門, 助手 (10177058) 仙波憲太郎東京大学医科学研究所, 制癌研究部, 助手 (70206663) 小安重夫 (財)東京都臨床医学総合研究所, 細胞生物学研究部 (90153684)
キーワード	癌 / 癌遺伝子 / リン酸化 / 蛋白質リン酸化酵素 / 癌抑制遺伝子 / シグナル伝達 / 細胞増殖 / チロシンキナ-ゼ
研究概要	1.estrogen(E2)がErbBー2のチロシンキナ-ゼ(PTK)活性を活性化しdown regulationをひきおこすことを見出した。2.B細胞抗原受容体を刺激することによりLynが活性化されること、Fyn過剰発現によりT細胞受容体を介した情報伝達反応が促進されることが示された。3.cーSrcのTyrー527をリン酸化化するPTK(CSK)のcDNAクロ-ニングに成功した。4.Crk蛋白質がリン酸化チロシンを有する蛋白質と特異的に結合することを示した。5.ラット肝からのAJの単離精製に成功し、Src,Yes,Lyn蛋白質が局在していることを見出し、さらにYes蛋白質と複合体を形成する分子の候補を同定した。またAJのリン酸化チロシンが細胞周期により変動するという結果を得た。6.MAPキナ-ゼの完全精製と、cDNAクロ-ニングに成功した。またMAPキナ-ゼがG0/G1期だけでなくS期およびM期にも活性化し細胞質のみでなく核にも局在する事を明らかにした。Rafー1キナ-ゼについては、活性化型は正常化型と比べ膜や核への局在が多いこと、増殖刺激や形質転換によってリン酸化が増大することを見出した。新しい遺伝子検索細胞SHOKを用いて単離したcot遺伝子が新しいタイプのSーG2期に活性化するPS/TKをコ-ドしていることを明らかにした。またヒトcーcot遺伝子を単離、構造決定し産物を同定した。7.Jun結合蛋白質のリン酸化状態が形質転換に伴い、顕著に変化することを見い出した。cdk2キナ-ゼおよびMAPキナ-ゼがRBキナ-ゼである可能性を示唆する実験結果を得た。またRBと結合する細胞蛋白質をコ-ドするcDNAのクロ-ニングに成功した。8.高品質cDNAライブラリ-の作製、大腸菌、酵母への効率の良い遺伝子導入法を開発し酵母の細胞周期変異を相補するヒト遺伝子(weel,cdc25)を単離した。

研究成果
(36件)

すべてその他

すべて文献書誌 (36件)

[文献書誌] Matsuda,M.: "Identifiction of domains of the v-crk on cogene product sufficient for association with phosphotyrosine-containing." Mol,Cell.Biol. 11. 1607-1613 (1991)
[文献書誌] Matsuda,M.: "Biological and biochemical activity of v-Crk chimeras containing the SH_2/SH_3 regions of PLC-r and Src." J.Virol.66. 115-121 (1992)
[文献書誌] Takahashi,S: "Thyrotropin potentiation of insulin-like growth factor-I depentein protein synthesis in FRTL-5 cells" Endocrinology.
[文献書誌] Takahashi,S: "Thyrotropin induction of tyrosine kinases in FRTL-5 cells" Endocrinology.
[文献書誌] Kazushi,I.: "Human c-fgr induces a monocyte-speafic enzyme in NIH3T3 cells." Mol.Cell.Biol.11. 6279-6285 (1991)
[文献書誌] Okada,M.: "CSK:a protein-tyrosein kinase involved in regulation of src family kinases." J.Biol.Chem.266. 24249-24252 (1991)
[文献書誌] Kazuyoshi,A: "Purification and Characterization of a Rat Liver Membrane Tyrosine-Protein kinase,the Possible Protooncogene c-yes Product,p60c-yes." J.Biol.Chem.266. 4831-4839 (1991)
[文献書誌] Sato,N.: "Radixin,a barbed-end-capping acti-modulating protein,is concentrated at the cloavage furrow during cytoknesis." J.Cell.Biol.113. 321-330 (1991)
[文献書誌] Tsukita,Sa: "Speific proto-oncogenic tyrosine kinases of src family arc entichecl in cell-to-cell adhorens junctions,where the level of tyrosine phosphorylation is elevated." J.Cell.Biol.113. 867-879 (1991)
[文献書誌] Funayama,N: "Radixin is a novel member of the band 4.1 family." J.Cell.Biol. 115. 1039-1048 (1991)
[文献書誌] Itoh,M: "A 220KD undercoat-constitutive protein:Its specific localization at cadherin-based cell-to-cell adhesion sves." J.Cell.Biol. 115. 1449-1462 (1991)
[文献書誌] Akiyama,T.: "Use and specificity of genistein as an inhibitor of protein tyrosine kinase" Methods in Enzymology. 201. 362-370 (1991)
[文献書誌] Iwamoto,T: "Preferntial development of pre-B lymphoma with drastically down-regulated N-myc in the E_μ-ret transgenic mice." Eur.J.Immunol. 21. 1809-1814 (1991)
[文献書誌] Ishizaka,Y: "A high phosphorylation state and increased activity of the tre motif in the NIH3T3 cell transformant induced by ret TPC." Biochem.Biophys.Res.Commun.179. 1331-1336 (1991)
[文献書誌] Iwamoto,T: "Aberrant melanogenesis and melanocytic tamor development in transgenic mice that carry metallothiorein/ret fusion gene." EMPO.J.10. 3167-3175 (1991)
[文献書誌] Taniguchi,M.: "The ret on cogene products are membrance-bound glycoproteins phosphorylated on tyrosine residues in vivo." Biochem.Biophys.Res.Commun.181. 416-422 (1991)
[文献書誌] Takahashi,M.: "Proliferation and neoplastic transformation of pigment cells in metallothionein/ret transgenic mice." Pigment Cell Res.
[文献書誌] Iwamoto,T.: "The ret oncogene can incluce melanogenesis and melanocyte development in W^v/W^v mice." Exp.Cell.Res.
[文献書誌] A,Masuda: "Signal transduction cascade shard by epidermal growth factor is a major pathway for oncogenic transformation in NRR cells." The New Biologist. (1992)
[文献書誌] Kume,K.: "On cogenic signal-induced ability to enter S phase in the abence of anchorage is the mechanism for the growth of trans formed NRK,cells in soft agar." The New Biologist. (1992)
[文献書誌] Osawa,M: "Histological factors contributing to favorable prognoses of soft tissue sarcomas in premenopausal patients." Jpn.J.Clin.Oncol. 21. 115-119 (1991)
[文献書誌] Fujita,M.Q.: "Structure of the dnaA and DnaA-box region in Mycoplasma capricolm chromosome:conservation and variations in the course of evolution." Gene. (1992)
[文献書誌] Fujita,M.Q: "Maintenance of androgen-,glucocorticoid-or estrogen-responsive growth in shionogi carcinoma subline sustained in castrated mice with high does of estrogen for 30 generations (3years)" Submitted to Jpn.J.Cancer Res.
[文献書誌] Takahashi,M.: "Identification of the ret proto-oncogene prducts in neuroblastoma and leulcemia cells." Oncogene. 6. 297-301 (1991)
[文献書誌] Nada,S.: "Cloning of a complementary DNA for a protein-tyrosine kinase that specifically phosphorylates a negative regulatory site of p60^<c-src>." Nature. 351. 69-72 (1991)
[文献書誌] Gotoh,Y.: "In vitro effects on microtuble dynamics of purified Xenopus M phase-activated MAP kinase." Nature. 349. 251-254 (1991)
[文献書誌] Gotoh,Y: "Xenopus M phase MAP kinase:isolation of its cDNA and activation by MPF." EMBO J.10. 2661-2668 (1991)
[文献書誌] Hoshi,M: "MAP kinase-catalyzed phosphorylation of microtubuleassociated proteins,MAP2 and MAP4,induces their fanctional alteration." Eur.J.Biochem.
[文献書誌] Matsuda,S: "Xenopus MAP kinase activator:identification and function as a key intermediate in the phosphorylation cascade." EMBO J.
[文献書誌] Nishida,E.: "MAP kinase and cytoskeleton in the mitogenic signal transduction." Int.Rev.Cytol.
[文献書誌] Akiyama,T.: "The transforming protential of the c-erbB-2 protein is regulated by its autophosphorylation at the carboxylterminal domain." Mol.Cell.Biol. 11. 833-842 (1991)
[文献書誌] Saga,T.: "Scintigraphic detection of overexpressed c-erbB-2 Proto-onoogene product by a class-switched murine anti-c-erbB-2 protein nonoclonal antibody." Cancer Res.51. 990-994 (1991)
[文献書誌] Fujimoto,A: "Transactivation of the TPA-responsive element by the oncogenic c-erbB-2 protein is partly mediated by protein kinase C." Biochem.Biophys.Res.Commun.178. 724-734 (1991)
[文献書誌] Aoki,M: "Identification and characterization of protein products of the cot oncogene with serine kinase activity." Oncogene. 6. 1515-1520 (1991)
[文献書誌] Moriyama,M.: "Expression of c-erbB-2 gene product in urinary bladder cancer" J.Urology. 145. 423-427 (1991)
[文献書誌] 豊島久真男: "癌化と増殖のシグナル伝達" 羊土社, 236 (1991)

1991 年度 実績報告書

細胞癌化における蛋白質リン酸化

研究代表者

秋山 徹 微生物病研究所, 発癌遺伝子部門, 助教授 (70150745)

研究成果

[文献書誌] Matsuda,M.: "Identifiction of domains of the v-crk on cogene product sufficient for association with phosphotyrosine-containing." Mol,Cell.Biol. 11. 1607-1613 (1991)

[文献書誌] Matsuda,M.: "Biological and biochemical activity of v-Crk chimeras containing the SH_2/SH_3 regions of PLC-r and Src." J.Virol.66. 115-121 (1992)

[文献書誌] Takahashi,S: "Thyrotropin potentiation of insulin-like growth factor-I depentein protein synthesis in FRTL-5 cells" Endocrinology.

[文献書誌] Takahashi,S: "Thyrotropin induction of tyrosine kinases in FRTL-5 cells" Endocrinology.

[文献書誌] Kazushi,I.: "Human c-fgr induces a monocyte-speafic enzyme in NIH3T3 cells." Mol.Cell.Biol.11. 6279-6285 (1991)

[文献書誌] Okada,M.: "CSK:a protein-tyrosein kinase involved in regulation of src family kinases." J.Biol.Chem.266. 24249-24252 (1991)

[文献書誌] Kazuyoshi,A: "Purification and Characterization of a Rat Liver Membrane Tyrosine-Protein kinase,the Possible Protooncogene c-yes Product,p60c-yes." J.Biol.Chem.266. 4831-4839 (1991)

[文献書誌] Sato,N.: "Radixin,a barbed-end-capping acti-modulating protein,is concentrated at the cloavage furrow during cytoknesis." J.Cell.Biol.113. 321-330 (1991)

[文献書誌] Tsukita,Sa: "Speific proto-oncogenic tyrosine kinases of src family arc entichecl in cell-to-cell adhorens junctions,where the level of tyrosine phosphorylation is elevated." J.Cell.Biol.113. 867-879 (1991)

[文献書誌] Funayama,N: "Radixin is a novel member of the band 4.1 family." J.Cell.Biol. 115. 1039-1048 (1991)

[文献書誌] Itoh,M: "A 220KD undercoat-constitutive protein:Its specific localization at cadherin-based cell-to-cell adhesion sves." J.Cell.Biol. 115. 1449-1462 (1991)

[文献書誌] Akiyama,T.: "Use and specificity of genistein as an inhibitor of protein tyrosine kinase" Methods in Enzymology. 201. 362-370 (1991)

[文献書誌] Iwamoto,T: "Preferntial development of pre-B lymphoma with drastically down-regulated N-myc in the E_μ-ret transgenic mice." Eur.J.Immunol. 21. 1809-1814 (1991)

[文献書誌] Ishizaka,Y: "A high phosphorylation state and increased activity of the tre motif in the NIH3T3 cell transformant induced by ret TPC." Biochem.Biophys.Res.Commun.179. 1331-1336 (1991)

[文献書誌] Iwamoto,T: "Aberrant melanogenesis and melanocytic tamor development in transgenic mice that carry metallothiorein/ret fusion gene." EMPO.J.10. 3167-3175 (1991)

[文献書誌] Taniguchi,M.: "The ret on cogene products are membrance-bound glycoproteins phosphorylated on tyrosine residues in vivo." Biochem.Biophys.Res.Commun.181. 416-422 (1991)

[文献書誌] Takahashi,M.: "Proliferation and neoplastic transformation of pigment cells in metallothionein/ret transgenic mice." Pigment Cell Res.

[文献書誌] Iwamoto,T.: "The ret oncogene can incluce melanogenesis and melanocyte development in W^v/W^v mice." Exp.Cell.Res.

[文献書誌] A,Masuda: "Signal transduction cascade shard by epidermal growth factor is a major pathway for oncogenic transformation in NRR cells." The New Biologist. (1992)

[文献書誌] Kume,K.: "On cogenic signal-induced ability to enter S phase in the abence of anchorage is the mechanism for the growth of trans formed NRK,cells in soft agar." The New Biologist. (1992)

[文献書誌] Osawa,M: "Histological factors contributing to favorable prognoses of soft tissue sarcomas in premenopausal patients." Jpn.J.Clin.Oncol. 21. 115-119 (1991)

[文献書誌] Fujita,M.Q.: "Structure of the dnaA and DnaA-box region in Mycoplasma capricolm chromosome:conservation and variations in the course of evolution." Gene. (1992)

[文献書誌] Fujita,M.Q: "Maintenance of androgen-,glucocorticoid-or estrogen-responsive growth in shionogi carcinoma subline sustained in castrated mice with high does of estrogen for 30 generations (3years)" Submitted to Jpn.J.Cancer Res.

[文献書誌] Takahashi,M.: "Identification of the ret proto-oncogene prducts in neuroblastoma and leulcemia cells." Oncogene. 6. 297-301 (1991)

[文献書誌] Nada,S.: "Cloning of a complementary DNA for a protein-tyrosine kinase that specifically phosphorylates a negative regulatory site of p60^<c-src>." Nature. 351. 69-72 (1991)

[文献書誌] Gotoh,Y.: "In vitro effects on microtuble dynamics of purified Xenopus M phase-activated MAP kinase." Nature. 349. 251-254 (1991)

[文献書誌] Gotoh,Y: "Xenopus M phase MAP kinase:isolation of its cDNA and activation by MPF." EMBO J.10. 2661-2668 (1991)

[文献書誌] Hoshi,M: "MAP kinase-catalyzed phosphorylation of microtubuleassociated proteins,MAP2 and MAP4,induces their fanctional alteration." Eur.J.Biochem.

[文献書誌] Matsuda,S: "Xenopus MAP kinase activator:identification and function as a key intermediate in the phosphorylation cascade." EMBO J.

[文献書誌] Nishida,E.: "MAP kinase and cytoskeleton in the mitogenic signal transduction." Int.Rev.Cytol.

[文献書誌] Akiyama,T.: "The transforming protential of the c-erbB-2 protein is regulated by its autophosphorylation at the carboxylterminal domain." Mol.Cell.Biol. 11. 833-842 (1991)

[文献書誌] Saga,T.: "Scintigraphic detection of overexpressed c-erbB-2 Proto-onoogene product by a class-switched murine anti-c-erbB-2 protein nonoclonal antibody." Cancer Res.51. 990-994 (1991)

[文献書誌] Fujimoto,A: "Transactivation of the TPA-responsive element by the oncogenic c-erbB-2 protein is partly mediated by protein kinase C." Biochem.Biophys.Res.Commun.178. 724-734 (1991)

[文献書誌] Aoki,M: "Identification and characterization of protein products of the cot oncogene with serine kinase activity." Oncogene. 6. 1515-1520 (1991)

[文献書誌] Moriyama,M.: "Expression of c-erbB-2 gene product in urinary bladder cancer" J.Urology. 145. 423-427 (1991)

[文献書誌] 豊島 久真男: "癌化と増殖のシグナル伝達" 羊土社, 236 (1991)

1991 年度実績報告書

秋山徹微生物病研究所, 発癌遺伝子部門, 助教授 (70150745)

[文献書誌] 豊島久真男: "癌化と増殖のシグナル伝達" 羊土社, 236 (1991)