1994 年度実績報告書

細胞増殖・癌化のシグナル伝達とリン脂質代謝

研究課題

研究課題/領域番号	06281108
研究機関	岐阜大学
研究代表者	岡野幸雄岐阜大学, 医学部, 教授 (10177066)
研究分担者	福井泰久東京大学, 農学院, 助教授 (00181248) 遠藤剛千葉大学, 理学部, 助教授 (30194038) 浜口道成名古屋大学, 医学部, 教授 (90135351) 稲垣昌樹東京都老人研究所, 部門長 (30183007) 貝淵弘三奈良先端科学技術大学院大学, 教授 (00169377)
キーワード	ホスホリパーゼD / ホスホリパーゼC / src / PI-3キナーゼ / 細胞骨格蛋白質 / SH2 / SH3 / Cキナーゼ / MAPキナーゼ
研究概要	1.ras細胞のCa^<2+>オシレーションに関与するカチオンチャネルは、イノシトールポリリン酸特に、IP4(3,4,5,6)によって開口されることが示唆された。 2.ras細胞やPAF受容体高発現CHO細胞における刺激に伴うPLDの活性化にPLCγ1やある種のチロシンキナーゼが関与することが示唆された。 3.マウス胎児繊維芽細胞(MEF)の継代培養の過程で消失する新規の細胞蛋白質p33を発現した。p33の抗体は不死化細胞のG1/S移行を阻止した。 4.PLCγSH2/SH3領域およびアンチセンスAshの細胞内導入は、c-junを介したアポトーシスを誘導し、dominant negative分子としての機能が示唆された。 5.PI-3Kの特異的な阻害剤であるwortmanninを用いて、PI-3Kが破骨細胞における膜融合やアクチンの構造形成に必須であることが明らかとなった。 6.α-アクチニンのアミノ酸168-184がPIP2結合部位であり、部分精製PLCの阻害作用から、その塩基性アミノ酸の重要性を証明した。 7.p75^<HS1>のLynとSykによる2つの独占的チロシンリン酸化部位を同定し、細胞質から核への移行を示した。 8.srcの細胞癌化能とMMP分泌亢進能の一致を示し、srcによるガドヘリン機能の阻害がカテニン構造の差に由来しないことを明らかにした。 9.化学発癌大腸腫瘍の近傍大腸粘膜のPLC-δ1の発現低下を認めた。しかし、発癌物質処置粘膜と無処置粘膜との間に有意な差は認めなかった。 10.表皮顆粒層で強く発現するnPKCηを活性化するコレステロール硫酸は、化学発癌による皮膚腫瘍の発生を顕著に抑制した。 11.PGF2α刺激グリア細胞のビメンチン部位特異的リン酸化反応はおもにCa^<2+>-カルモジュリン系によって担われていることが明らかとなった。 12.ツメガエル卵とウシ脳からREKSを精製し、100kDaのキナーゼであること、14-3-3蛋白質によって活性化されることを明らかにした。 13.MAPKKKのアミノ酸配列をもとにMUKI1〜3の3種の新規キナーゼを単離し、MUK1を介する哺乳類の新しいシグナル系の存在が示唆された。 14.カルパイン高発現細胞株およびカルパイン自己消化中間体の抗体を用いて、カルパインによるアクチンフィラメントの制御を明らかにした。 15.変異ゲルゾリンはアクチン重合核形成能の増大とアクチン繊維切断能の低下をきたし、細胞内発現は増殖遅延をもたらした。

研究成果
(28件)

すべてその他

すべて文献書誌 (28件)

[文献書誌] Banno,Y.: "Thrombin-mediated phosphoinositide hydrolysis in chines hamster ovary cells overexpressing phospholipase C-δ1." J.Biol.Chem.269. 15846-15852 (1994)
[文献書誌] Alam,M.S.: "Defective phospholipase D(PLD) activation in Ki-ras-transformed NIH-3T3 cells:evidence for downstream effector of PLCγl in PDGF-mediated signal transduction." Biochem.Biophys.Res.Commun.(in press).
[文献書誌] Liu,B.: "Activation of phospholipase D in Chinese hamster ovary cells expressing platelet-activating factor receptor." J.Biochem.116,. 882-891 (1993)
[文献書誌] Wadhwa,R.: "Spontaneous immortalization of mouse fibroblasts involves structural chanfes in senescence inducing protein,mortalin." Biochem.Biophys.Res.Commun.197. 202-206 (1993)
[文献書誌] Wadhwa,R.: "Correlation between complementation group for immortality and the cellular distribution of mortalin." Expl.Cell Res.216. 101-106 (1995)
[文献書誌] Fukami,K.: "α-Actinin and vinculin are PIP2-binding proteins involved in signaling by tyrosine kinase." J.Biol.Chem.269. 1518-1522 (1994)
[文献書誌] Ohkubo,Y.: "SV40 large T antigen reinduces the cell cycle in terminally differentiated myotubes through inducing Cdk2,Cdc2,and their partner cyclins." Expl.Cell.Res.214. 270-278 (1994)
[文献書誌] Kimura,K.: "Neurite outgrowth of PC12 cells is suppressed by wortmannin,a specific inhibitor of phosphatidylinositol-3 kinase." J.Biol.Chem.269. 18961-18967 (1994)
[文献書誌] Daduang,S.: "Production of monoclonal antibodies specific to carboxyl terminal region of 85kDa subunit of phosphatidylinositol 3-kinase." Immun.Cell Biol.(in press).
[文献書誌] Hay,J.C.: "ATP-dependent inositide phosphorylation required for Ca^<2+>-activated Seceretion." Nature. (in press).
[文献書誌] Fukami,K.: "α-Actinin and vinculin are PIP2-binding proteins involved in signaling by tyrosine kinase." J.Biol.Chem.269. 1518-1522 (1994)
[文献書誌] Kurosaki,T.: "Syk activation by the Src-family tyrosine kinase in the B-cell receptor signaling." J.Exp.Med.17. 1725-1729 (1994)
[文献書誌] Hamaguchi,M.: "Augmentation of metalloproteinase(gelatinase)activity secreted from Rous sarcoma virus-infected cel correlates with transforming activity of src." Oncogene. (in press).
[文献書誌] Iwata,H.: "Abundant but inactive-state gp140 proto-trk is expressed in neuroblastoma of patients with good prognosis." Jpn.J.Cancer Res.85. 32-39 (1994)
[文献書誌] Yoshimi,N.: "Reduced expression of phospholipase C-δ,a signal-tranducing enzyme,in rat colon neoplasms induced by methylazoxymethanol acetate." Mol.Carcinogenesis,. 11. 192-196 (1994)
[文献書誌] Mori,H.: "Chemopreventative effects of plant derived phenolic organosulfur and other compounds on carcinogenesis in digestive organs." Environ.Mut.Res.Commun.(in press).
[文献書誌] Chida,K.: "The η isoform of protein kinase C is localized on rough endoplasmic reticulum." Mol.Cell.Biol.14. 3782-3790 (1994)
[文献書誌] Ikuta,T.: "Cholesterol sulfate,a novel activator for the η isoform of protein kinase C." Cell Growth & Differentiation. 5. 943-947 (1994)
[文献書誌] Saido,T.C.: "Calpain:new perspectives in molecular diversity and physiological-pathological involvement." FASEB J.8. 814-822 (1994)
[文献書誌] Saito,Y.: "The calpain-calpastatin system is regulated differentially during human neuroblastoma cell differentiation to Schwannian and neuronal cells." FEBS Lett.353. 327-331 (1994)
[文献書誌] Fujita,H.: "Functions of mutant gelsolin His321 isolated from a flat revertant of Ras-transformed cells." Eur.J.Biochem.(in press).
[文献書誌] Ishizaki,A.: "Growth-inhibitory functions of a mutated gelsolin (His321) in NIH/3T3 mouse fibroblasts." Expl.Cell.Res.(in press).
[文献書誌] Tsujimura,K.: "Visualization and function of vimentin phosphorylation by sds2 kinase during mitosis." J.Biol.Chem.269. 31097-31106 (1994)
[文献書誌] Inagaki,N.: "Spatiotemporal distribution of protein kinase and phosphatase activities." Trends Biochem.Sci.19. 448-452 (1994)
[文献書誌] Nakanishi,H.: "Different functions of smg GDP dissociation stimulator and mammalian counterpart of yeast Cdc25." J.Biol.Chem.269. 15085-15091 (1994)
[文献書誌] Shimuzu,K.: "Synergistic activation by Ras and 14-3-3 protein of MAP kinase kinase kinase nemed Ras-dependent ERK kinase stimulator(REKS)." J.Biol.Chem.269. 22917-22920 (1994)
[文献書誌] Osada,S-i.: "MUK3,a protein kinase related to STE11,Byr2,raf,and mos-MAPKKK family and MLK/SPRK/PTK protein kinases,with putative leucine-zipper and SH3-binding motifs." Biochem.Biophys.Res.Commun.(in press).
[文献書誌] Hirano,M.: "A protein kinase C isozyme,nPKCe,is involved in the activation of NF-kB by 12-0-tetradecanoylphorbol 13-acetate(TPA) in rat 3Y1 fibroblasts." Biochem.Biophys.Res.Commun.206. 429-436 (1995)

1994 年度 実績報告書

細胞増殖・癌化のシグナル伝達とリン脂質代謝

研究代表者

岡野 幸雄 岐阜大学, 医学部, 教授 (10177066)

研究成果

[文献書誌] Banno,Y.: "Thrombin-mediated phosphoinositide hydrolysis in chines hamster ovary cells overexpressing phospholipase C-δ1." J.Biol.Chem.269. 15846-15852 (1994)

[文献書誌] Alam,M.S.: "Defective phospholipase D(PLD) activation in Ki-ras-transformed NIH-3T3 cells:evidence for downstream effector of PLCγl in PDGF-mediated signal transduction." Biochem.Biophys.Res.Commun.(in press).

[文献書誌] Liu,B.: "Activation of phospholipase D in Chinese hamster ovary cells expressing platelet-activating factor receptor." J.Biochem.116,. 882-891 (1993)

[文献書誌] Wadhwa,R.: "Spontaneous immortalization of mouse fibroblasts involves structural chanfes in senescence inducing protein,mortalin." Biochem.Biophys.Res.Commun.197. 202-206 (1993)

[文献書誌] Wadhwa,R.: "Correlation between complementation group for immortality and the cellular distribution of mortalin." Expl.Cell Res.216. 101-106 (1995)

[文献書誌] Fukami,K.: "α-Actinin and vinculin are PIP2-binding proteins involved in signaling by tyrosine kinase." J.Biol.Chem.269. 1518-1522 (1994)

[文献書誌] Ohkubo,Y.: "SV40 large T antigen reinduces the cell cycle in terminally differentiated myotubes through inducing Cdk2,Cdc2,and their partner cyclins." Expl.Cell.Res.214. 270-278 (1994)

[文献書誌] Kimura,K.: "Neurite outgrowth of PC12 cells is suppressed by wortmannin,a specific inhibitor of phosphatidylinositol-3 kinase." J.Biol.Chem.269. 18961-18967 (1994)

[文献書誌] Daduang,S.: "Production of monoclonal antibodies specific to carboxyl terminal region of 85kDa subunit of phosphatidylinositol 3-kinase." Immun.Cell Biol.(in press).

[文献書誌] Hay,J.C.: "ATP-dependent inositide phosphorylation required for Ca^<2+>-activated Seceretion." Nature. (in press).

[文献書誌] Fukami,K.: "α-Actinin and vinculin are PIP2-binding proteins involved in signaling by tyrosine kinase." J.Biol.Chem.269. 1518-1522 (1994)

[文献書誌] Kurosaki,T.: "Syk activation by the Src-family tyrosine kinase in the B-cell receptor signaling." J.Exp.Med.17. 1725-1729 (1994)

[文献書誌] Hamaguchi,M.: "Augmentation of metalloproteinase(gelatinase)activity secreted from Rous sarcoma virus-infected cel correlates with transforming activity of src." Oncogene. (in press).

[文献書誌] Iwata,H.: "Abundant but inactive-state gp140 proto-trk is expressed in neuroblastoma of patients with good prognosis." Jpn.J.Cancer Res.85. 32-39 (1994)

[文献書誌] Yoshimi,N.: "Reduced expression of phospholipase C-δ,a signal-tranducing enzyme,in rat colon neoplasms induced by methylazoxymethanol acetate." Mol.Carcinogenesis,. 11. 192-196 (1994)

[文献書誌] Mori,H.: "Chemopreventative effects of plant derived phenolic organosulfur and other compounds on carcinogenesis in digestive organs." Environ.Mut.Res.Commun.(in press).

[文献書誌] Chida,K.: "The η isoform of protein kinase C is localized on rough endoplasmic reticulum." Mol.Cell.Biol.14. 3782-3790 (1994)

[文献書誌] Ikuta,T.: "Cholesterol sulfate,a novel activator for the η isoform of protein kinase C." Cell Growth & Differentiation. 5. 943-947 (1994)

[文献書誌] Saido,T.C.: "Calpain:new perspectives in molecular diversity and physiological-pathological involvement." FASEB J.8. 814-822 (1994)

[文献書誌] Saito,Y.: "The calpain-calpastatin system is regulated differentially during human neuroblastoma cell differentiation to Schwannian and neuronal cells." FEBS Lett.353. 327-331 (1994)

[文献書誌] Fujita,H.: "Functions of mutant gelsolin His321 isolated from a flat revertant of Ras-transformed cells." Eur.J.Biochem.(in press).

[文献書誌] Ishizaki,A.: "Growth-inhibitory functions of a mutated gelsolin (His321) in NIH/3T3 mouse fibroblasts." Expl.Cell.Res.(in press).

[文献書誌] Tsujimura,K.: "Visualization and function of vimentin phosphorylation by sds2 kinase during mitosis." J.Biol.Chem.269. 31097-31106 (1994)

[文献書誌] Inagaki,N.: "Spatiotemporal distribution of protein kinase and phosphatase activities." Trends Biochem.Sci.19. 448-452 (1994)

[文献書誌] Nakanishi,H.: "Different functions of smg GDP dissociation stimulator and mammalian counterpart of yeast Cdc25." J.Biol.Chem.269. 15085-15091 (1994)

[文献書誌] Shimuzu,K.: "Synergistic activation by Ras and 14-3-3 protein of MAP kinase kinase kinase nemed Ras-dependent ERK kinase stimulator(REKS)." J.Biol.Chem.269. 22917-22920 (1994)

[文献書誌] Osada,S-i.: "MUK3,a protein kinase related to STE11,Byr2,raf,and mos-MAPKKK family and MLK/SPRK/PTK protein kinases,with putative leucine-zipper and SH3-binding motifs." Biochem.Biophys.Res.Commun.(in press).

[文献書誌] Hirano,M.: "A protein kinase C isozyme,nPKCe,is involved in the activation of NF-kB by 12-0-tetradecanoylphorbol 13-acetate(TPA) in rat 3Y1 fibroblasts." Biochem.Biophys.Res.Commun.206. 429-436 (1995)

1994 年度実績報告書

岡野幸雄岐阜大学, 医学部, 教授 (10177066)