1991 年度実績報告書

細胞内情報伝達系と細胞増殖の制御機構

研究課題

研究課題/領域番号	03258229
研究機関	神戸大学
研究代表者	高井義美神戸大学, 医学部, 教授 (60093514)
研究分担者	河田正仁神戸大学, 医学部, 助手 (20224785) 菊池章神戸大学, 医学部, 助手 (10204827) 貝淵弘三神戸大学, 医学部, 助教授 (00169377)
キーワード	細胞増殖 / ras p21 / 低分子量GTP結合蛋白質 / smg GDS / rafキナ-ゼ / 'cーfos遺伝子
研究概要	最近の私共や他の研究室の解析により、ras p21が細胞増殖因子のトランスデュ-サ-として働き、rafキナ-ゼを介して情報を伝達している可能性がある。本年度はras p21とrafキナ-ゼの活性化機構と作用点に焦点を当てて解析を行い、以下の結果を得た。私共はすでにras p21類似低分子量GTP結合蛋白質であるsmg p21のGDP/GTP交換反応を促進する蛋白質、smg GDSを精製してそのcDNAをクロ-ニングしている。ras p21のGDP/GTP交換反応を制御する蛋白質は現在まで同定されていなかったが、本年度、私共はsmg GDSがsmg p21のみならず、Kiーras p21にも同様に作用することを明らかにした。また、smg GDSの作用にこれらの低分子量GTP結合蛋白質の翻訳後修飾が不可欠であることも明らかにした。さらに、NIH/3T3細胞においてsmg GDSがKiーras p21と共同してcーfos遺伝子の発現を促進すると共に、細胞を形質転換させることを明らかにした。また、人癌におけるsmg GDS遺伝子の変異を検索するために、ヒトsmg GDSのcDNAをクロ-ニングした。一方、smg p21はras p21のエフェクタ-領域と同じアミノ酸配列を有しており、ras p21の作用に拮抗するか同様の作用を示すと推定されている。私共はsmg p21がPDGF、Cキナ-ゼやras p21によるcーfos遺伝子の発現を抑制するが、Aキナ-ゼやrafキナ-ゼによるcーfos遺伝子の発現を抑制しないことを明らかにした。本年度の私共の研究成果から、ras p21のGDP/GTP交換反応を調節する蛋白質の性状が明らかとなったと共に、PDGF受容体とCキナ-ゼの下流にras p21が存在し、さらにその下流にrafキナ-ゼが存在することがほぼ確実になった。以上の成積を得たことにより本年度の研究計画はほぼ達成することができた。

研究成果
(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] Hiroyoshi,M.: "Role of the Cーterminal region of smg p21,a ras p21ーlike small GTPーbinding protein,in membrane and smg p21 GDP/GTP exchange protein interactions." J.Biol.Chem.266. 2962-2969 (1991)
[文献書誌] Hata,Y.: "Enhancement of the actions of smg p21 GDP/GTP exchange protein by the protein kinase Aーcatalyzed phosphorylation of smg p21." J.Biol.Chem.266. 6571-6577 (1991)
[文献書誌] Katayama,M.: "The posttranslationally modified Cーterminal structure of bovine aortic smooth muscle rhoA p21." J.Biol.Chem.266. 12639-12645 (1991)
[文献書誌] Horiuchi,H.: "A novel prenyltransferase for a small GTPーbinding protein having a Cーterminal CysーAlaーCys structure." J.Biol.Chem.266. 16981-16984 (1991)
[文献書誌] Shirataki,H.: "Inhibition of the action of the stimulatory GDP/GTP exchange protein for smg p21 by the geranylgeranylated synthetic peptides designed from its Cーterminal region." J.Biol.Chem.266. 20672-20677 (1991)
[文献書誌] Araki,S.: "Role of the Cーterminal region of smg p25A in its interaction with membranes and the GDP/GTP exchange protein." Mol.Cell.Biol.11. 1438-1447 (1991)
[文献書誌] Kaibuchi,K.: "Molecular cloning of the cDNA for stimulatory GDP/GTP exchange protein for smg p21s(ras p21ーlike small GTPーbinding proteins)and characterization of stimulatory GDP/GTP exchange protein." Mol.Cell.Biol.11. 2873-2880 (1991)
[文献書誌] Sasaki,T.: "A mammalian inhibitory GDP/GTP exchange protein(GDP dissociation inhibitor)for smg p25A is active on the yeast SEC4 Protein." Mol.Cell.Biol.11. 2909-2912 (1991)
[文献書誌] Ueda,T.: "Purification and characterization from rat liver cytosol of a GDP dissociation inhibitor(GDI)for liver 24KG,a ras p21ーlike GTPーbinding protein,with properties similar to those of smg p25A GDI." Biochemistry. 30. 909-917 (1991)
[文献書誌] Farnsworth,C.C.: "C terminus of the small GTPーbinding protein smg p25A contains two geranylgeranylated cysteine residues and a methyl ester." Proc.Natl.Acod.Sci.USA. 88. 6196-6200 (1991)
[文献書誌] Mizuno,T.: "A stimulator GDP/GTP exchange proten for smg p21 is active on the postーtranslationally processed form of cーKiーras p21 and rhoA p21." proc.Natl.Acod.Sci.USA. 88. 6442-6446 (1991)
[文献書誌] Isomura,M.: "Regulation of binding of rhoB p20 to membranes by its specific regulatory protein,GDP dissociation inhibitor." Oncogene. 6. 119-124 (1991)
[文献書誌] Hori,Y.: "Postーtranslational modificationas of the Cーterminal region of the rho protein are important for its interaction with membranes and the stimulatory and inhibitory GDP/GTP exchange proteins." Oncogene. 6. 515-522 (1991)
[文献書誌] Kawata,M.: "The molecular heterogeneity of the smgー21/Krevー1/rap1 proteins,a GTPーbinding protein having the same effector domain as ras p21s,in bovine aortic smooth muscle membranes." Oncogene. 6. 841-848 (1991)
[文献書誌] Sano,K.: "Differential expression of the Nーmyc,cーfos,and smg p25A genes in human neuroblastoma cells during neuronal and Schwannian differentiation." Mol.Brain Res.9. 149-152 (1991)
[文献書誌] Takayama,K.: "Similarity of expression of low molecular weight G proteins smg p21A and ras p21 in normal and malignant human tissues." Cancer Res.51. 2223-2228 (1991)
[文献書誌] Kawamura,S.: "Inhibition of the action of a stimulatory GDP/GTP exchange protein for smg p21 by acidic membrane phospholipids." Jpn.J.Cancer Res.82. 758-761 (1991)
[文献書誌] Fujioka,H.: "Transforming and cーfos promoter/enhancerーstimulating activities of a stimulatory GDP/GTP exchange protein for small GTPーbinding proteins." J.Biol.Chem.267. 926-930 (1992)

1991 年度 実績報告書

細胞内情報伝達系と細胞増殖の制御機構

研究代表者

高井 義美 神戸大学, 医学部, 教授 (60093514)

研究成果

[文献書誌] Hiroyoshi,M.: "Role of the Cーterminal region of smg p21,a ras p21ーlike small GTPーbinding protein,in membrane and smg p21 GDP/GTP exchange protein interactions." J.Biol.Chem.266. 2962-2969 (1991)

[文献書誌] Hata,Y.: "Enhancement of the actions of smg p21 GDP/GTP exchange protein by the protein kinase Aーcatalyzed phosphorylation of smg p21." J.Biol.Chem.266. 6571-6577 (1991)

[文献書誌] Katayama,M.: "The posttranslationally modified Cーterminal structure of bovine aortic smooth muscle rhoA p21." J.Biol.Chem.266. 12639-12645 (1991)

[文献書誌] Horiuchi,H.: "A novel prenyltransferase for a small GTPーbinding protein having a Cーterminal CysーAlaーCys structure." J.Biol.Chem.266. 16981-16984 (1991)

[文献書誌] Shirataki,H.: "Inhibition of the action of the stimulatory GDP/GTP exchange protein for smg p21 by the geranylgeranylated synthetic peptides designed from its Cーterminal region." J.Biol.Chem.266. 20672-20677 (1991)

[文献書誌] Araki,S.: "Role of the Cーterminal region of smg p25A in its interaction with membranes and the GDP/GTP exchange protein." Mol.Cell.Biol.11. 1438-1447 (1991)

[文献書誌] Kaibuchi,K.: "Molecular cloning of the cDNA for stimulatory GDP/GTP exchange protein for smg p21s(ras p21ーlike small GTPーbinding proteins)and characterization of stimulatory GDP/GTP exchange protein." Mol.Cell.Biol.11. 2873-2880 (1991)

[文献書誌] Sasaki,T.: "A mammalian inhibitory GDP/GTP exchange protein(GDP dissociation inhibitor)for smg p25A is active on the yeast SEC4 Protein." Mol.Cell.Biol.11. 2909-2912 (1991)

[文献書誌] Ueda,T.: "Purification and characterization from rat liver cytosol of a GDP dissociation inhibitor(GDI)for liver 24KG,a ras p21ーlike GTPーbinding protein,with properties similar to those of smg p25A GDI." Biochemistry. 30. 909-917 (1991)

[文献書誌] Farnsworth,C.C.: "C terminus of the small GTPーbinding protein smg p25A contains two geranylgeranylated cysteine residues and a methyl ester." Proc.Natl.Acod.Sci.USA. 88. 6196-6200 (1991)

[文献書誌] Mizuno,T.: "A stimulator GDP/GTP exchange proten for smg p21 is active on the postーtranslationally processed form of cーKiーras p21 and rhoA p21." proc.Natl.Acod.Sci.USA. 88. 6442-6446 (1991)

[文献書誌] Isomura,M.: "Regulation of binding of rhoB p20 to membranes by its specific regulatory protein,GDP dissociation inhibitor." Oncogene. 6. 119-124 (1991)

[文献書誌] Hori,Y.: "Postーtranslational modificationas of the Cーterminal region of the rho protein are important for its interaction with membranes and the stimulatory and inhibitory GDP/GTP exchange proteins." Oncogene. 6. 515-522 (1991)

[文献書誌] Kawata,M.: "The molecular heterogeneity of the smgー21/Krevー1/rap1 proteins,a GTPーbinding protein having the same effector domain as ras p21s,in bovine aortic smooth muscle membranes." Oncogene. 6. 841-848 (1991)

[文献書誌] Sano,K.: "Differential expression of the Nーmyc,cーfos,and smg p25A genes in human neuroblastoma cells during neuronal and Schwannian differentiation." Mol.Brain Res.9. 149-152 (1991)

[文献書誌] Takayama,K.: "Similarity of expression of low molecular weight G proteins smg p21A and ras p21 in normal and malignant human tissues." Cancer Res.51. 2223-2228 (1991)

[文献書誌] Kawamura,S.: "Inhibition of the action of a stimulatory GDP/GTP exchange protein for smg p21 by acidic membrane phospholipids." Jpn.J.Cancer Res.82. 758-761 (1991)

[文献書誌] Fujioka,H.: "Transforming and cーfos promoter/enhancerーstimulating activities of a stimulatory GDP/GTP exchange protein for small GTPーbinding proteins." J.Biol.Chem.267. 926-930 (1992)

1991 年度実績報告書

高井義美神戸大学, 医学部, 教授 (60093514)