1989 Fiscal Year Annual Research Report

細胞分化の制御におけるCキナ-ゼとrasp21類似GTP結合蛋白質の作用機構

Research Project

Project/Area Number	01654512
Research Institution	Kobe University
Principal Investigator	高井義美神戸大学, 医学部, 教授 (60093514)
Co-Investigator(Kenkyū-buntansha)	溝口明神戸大学, 医学部, 講師 (90181916) 菊池章神戸大学, 医学部, 助手 (10204827) 貝淵弘三神戸大学, 医学部, 講師 (00169377)
Keywords	c-fos遺伝子 / Cキナ-ゼ / rasp21 / GTP結合蛋白質 / smgp21 / smgp25 / GAP / GDI
Research Abstract	種々の動物細胞の分化過程にc-fos遺伝子の発現が重要な役割を果たしていることが知られている。また、c-fos遺伝子の発現は、Cキナ-ゼやrasp21などの細胞内情報伝達系によって制御されていると考えられている。しかし、Cキナ-ゼや、c-fos遺伝子の分化過程における作用機構はなお解明されていない。私共は本研究で、構成的に活性化されるCキナ-ゼやAキナ-ゼのcDNAを構築して、活性型Cキナ-ゼやAキナ-ゼによるc-fos遺伝子の発現系を確立し、NIH/3T3細胞においてCキナ-ゼがSREを、Aキナ-ゼがCREを介してc-fos遺伝子の発現を促進することを明らかにした。また、rasp21やraf遺伝子産物はCキナ-ゼやAキナ-ゼとは非依存的にSREを介してc-fos遺伝子の発現を促進することも明らかにした。他方、私共はすでにrasp21に類似した新しいGTP結合蛋白質(G蛋白質)であるsmgp21とsmgp25の精製と構造決定に成功している。これらの、低分子量G蛋白質にはGDP結合型の不活性型とGTP結合型の活性型があり、不活性型と活性型の転換を制御する蛋白質が存在すると考えられる。rasp21にはそのGTPase活性を促進し、その結果GTP結合型とGDP結合型に転換する。GTPase活性促進蛋白質(GAP)が存在することが報告されているが、私共はrasp21に対するGAPを見出した。さらに、smgp25に対する新しい種類の活性調節蛋白質であるGDP解離抑制蛋白質(GDI)を均一蛋白質に精製し、そのcDNAを単離して一次構造を決定した。以上の結果、低分子量G蛋白質はGAPとGDIにより不活性型と活性型との転換が調節され、その活性が制御されていることが明らかとなった。今後、これらのG蛋白質やその制御蛋白質とCキナ-ゼなどの細胞内情報伝達系との関連についても解析を進めていく予定である。以上、本年度は予想以上の研究成果を達成することができた。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] Ohmori,T.: "Small molecular weight GTP-binding proteins in human platelet membranes Purification and characterization of a novel GTP-binding protein with a molecular weight of 22,000." J.Biol,Chem.264. 1877-1881 (1989)
[Publications] Kikuchi,A.: "Purification and characterization from bovine brain cytosol of two GTPase-activating proteins specific for smg p21,a GTP-binding protein having the same effector domain as c-ras p21s." J.Biol,Chem.264. 9133-9136 (1989)
[Publications] Kaibuchi,K.: "Molecular genetic analysis of the regulatory and catalytic domains of protein kinase C." J.Biol,Chem.264. 13489-13496 (1989)
[Publications] Kawata,M.: "Phosphorylation of smg p21,a ras p21-like GTP-binding protein,by cyclic AMP-dependent protein kinase in a cell-free system and in response to prostaglandin E_1 in intact human platelets." J.Biol,Chem.264. 15688-15695 (1989)
[Publications] Kaibuchi,K.: "Activation of the serum-response element and TPA-response element by the activated c-raf-1 protein in a manner independent of protein kinase C." J.Biol,Chem.264. 20855-20858 (1989)
[Publications] Adachi,M.: "Elevation of intracellular calcium ion by prostaglanding E_1 and its inhibition by protein kinase C in human megakaryocyte leukemia cell line." Cancer Res.49. 3805-3808 (1989)
[Publications] Mizoguchi,A.: "Localization and subcellular distribution of cellular ras gene products in rat brain." Mol.Brain Res.5. 31-44 (1989)
[Publications] Kim,S.: "Intrasynaptosomal distribution of the ras,rho and smg-25A GTP-binding proteins in bovine brain." Mol.Brain Res.6. 167-176 (1989)
[Publications] Sano,K.: "Tissue-specific expression of a novel GTP-binding protein(smg p25A)mRNA and its increase by nerve growth factor and cyclic AMP in rat pheochromocytoma PC-12 cells." Biochem.Biophys.Res.Commun.158. 377-385 (1989)
[Publications] Ueda,T.: "GTPase activating proteins for the smg-21 GTP-binding protein having the same effector domain as the ras proteins in human platelets." Biochem.Biophys.Res.Commun.159. 1411-1419 (1989)
[Publications] Kariya,K.: "Two types of protein kinase C with different functions in cultued rabbit aortic smooth muscle cells." Biochem.Biophys.Res.Commun.161. 1020-1027 (1989)
[Publications] Shoji,I.: "Kinetic analysis of the binding of guanine nucleotide to bovine brain smg p25A." Biochem.Biophys.Res.Commun.162. 273-281 (1989)
[Publications] Mizoguchi,A.: "Tissue distribution of smg p25A,a ras p21-like GTP-binding protein,studied by use of a specific monoclonal antibody." Biochem.Biophys.Res.Commun.162. 1438-1445 (1989)
[Publications] Kuroda,S.: "Kinetic analysis of the binding of guanine nucleotides to bovine brain rhoB p20,a ras p21-like GTP-binding protein" Biochem.Biophys.Res.Commun.163. 674-681 (1989)
[Publications] Kawata,M.: "Identification of a major GTP-binding protein in bovine aortic smooth muscle membranes as smg p21,a GTP-binding protein having the same effector domain as ras p21s." Biochem.Biophys.Res.Commun.162. 1418-1427 (1989)
[Publications] Ohga,N.: "Rabbit intestine contains a protein that ingibits the dissociation of GDP from and the subsequent binding fo GTP to rhoB p20,a ras p21-like GTP-binding protein." Biochem.Biophys.Res.Commun.163. 1523-1533 (1989)
[Publications] Yamamoto,K.: "Small molecular weight GTP-binding proteins and signal transduction." Clinica.Chimica.Acta.185. 347-356 (1989)
[Publications] Fukumoto,Y.: "Activation of the c-fos serum-response element by the activated c-Ha-ras protein in a manner independent of protein kinase C and cAMP-dependent protein kinase." J.Biol.Chem.265. 774-780 (1990)
[Publications] Sasaki,T.: "Purification and characterization from bovine brain cytosol of a protein that ingibits the dissociation of GDP from and the subsequent binding of GTP to smg p25A,a ras p21-like GTP-binding proein." J.Biol.Chem.265. 2333-2337 (1990)
[Publications] Hoshijima,M.: "Purification and characterization from bovine brain membranes of a GTP-binding protein with a Mr of 21,000 ADP-ribosylated by an ADP-ribosyltransferase contained in botulinum toxin type c1-Identification as the rhoA gene product." Mol.Brain Res.7. 9-16 (1990)
[Publications] Matsui,Y.: "Molecular cloning of smg p21B and identification of smg p21 purified from bovine brain and human platelets as smg p21B." Biochem.Biophys.Res.Commun.166. 1010-1016 (1990)
[Publications] Hata,Y.: "inhibition of the ras p21 GTPase-activating protein-stimulated GTPase activity of c-Ha-ras p21 by smg p21 gaving the same putative effector domain as ras p21s." J.Biol.Chem.(1990)
[Publications] Yamamoto,J.: "A GTPase-activating protein(GAP)for rhoB p20,a ras p21-like GTP-binding protein-Partial purification,characterization and subcellular distribution in rat brain." Mol.Brain Res.(1990)
[Publications] Kim,S.: "Tissue and subcellular distributions of smg p21s,ras p21-like GTP-binding proteins having the same putative effector domain as ras p21s,partly sistinct from those of c-ras p21s." Mol,Cell.Biol.(1990)

1989 Fiscal Year Annual Research Report

細胞分化の制御におけるCキナ-ゼとrasp21類似GTP結合蛋白質の作用機構

Principal Investigator

高井 義美 神戸大学, 医学部, 教授 (60093514)

Research Products

[Publications] Ohmori,T.: "Small molecular weight GTP-binding proteins in human platelet membranes Purification and characterization of a novel GTP-binding protein with a molecular weight of 22,000." J.Biol,Chem.264. 1877-1881 (1989)

[Publications] Kikuchi,A.: "Purification and characterization from bovine brain cytosol of two GTPase-activating proteins specific for smg p21,a GTP-binding protein having the same effector domain as c-ras p21s." J.Biol,Chem.264. 9133-9136 (1989)

[Publications] Kaibuchi,K.: "Molecular genetic analysis of the regulatory and catalytic domains of protein kinase C." J.Biol,Chem.264. 13489-13496 (1989)

[Publications] Kawata,M.: "Phosphorylation of smg p21,a ras p21-like GTP-binding protein,by cyclic AMP-dependent protein kinase in a cell-free system and in response to prostaglandin E_1 in intact human platelets." J.Biol,Chem.264. 15688-15695 (1989)

[Publications] Kaibuchi,K.: "Activation of the serum-response element and TPA-response element by the activated c-raf-1 protein in a manner independent of protein kinase C." J.Biol,Chem.264. 20855-20858 (1989)

[Publications] Adachi,M.: "Elevation of intracellular calcium ion by prostaglanding E_1 and its inhibition by protein kinase C in human megakaryocyte leukemia cell line." Cancer Res.49. 3805-3808 (1989)

[Publications] Mizoguchi,A.: "Localization and subcellular distribution of cellular ras gene products in rat brain." Mol.Brain Res.5. 31-44 (1989)

[Publications] Kim,S.: "Intrasynaptosomal distribution of the ras,rho and smg-25A GTP-binding proteins in bovine brain." Mol.Brain Res.6. 167-176 (1989)

[Publications] Sano,K.: "Tissue-specific expression of a novel GTP-binding protein(smg p25A)mRNA and its increase by nerve growth factor and cyclic AMP in rat pheochromocytoma PC-12 cells." Biochem.Biophys.Res.Commun.158. 377-385 (1989)

[Publications] Ueda,T.: "GTPase activating proteins for the smg-21 GTP-binding protein having the same effector domain as the ras proteins in human platelets." Biochem.Biophys.Res.Commun.159. 1411-1419 (1989)

[Publications] Kariya,K.: "Two types of protein kinase C with different functions in cultued rabbit aortic smooth muscle cells." Biochem.Biophys.Res.Commun.161. 1020-1027 (1989)

[Publications] Shoji,I.: "Kinetic analysis of the binding of guanine nucleotide to bovine brain smg p25A." Biochem.Biophys.Res.Commun.162. 273-281 (1989)

[Publications] Mizoguchi,A.: "Tissue distribution of smg p25A,a ras p21-like GTP-binding protein,studied by use of a specific monoclonal antibody." Biochem.Biophys.Res.Commun.162. 1438-1445 (1989)

[Publications] Kuroda,S.: "Kinetic analysis of the binding of guanine nucleotides to bovine brain rhoB p20,a ras p21-like GTP-binding protein" Biochem.Biophys.Res.Commun.163. 674-681 (1989)

[Publications] Kawata,M.: "Identification of a major GTP-binding protein in bovine aortic smooth muscle membranes as smg p21,a GTP-binding protein having the same effector domain as ras p21s." Biochem.Biophys.Res.Commun.162. 1418-1427 (1989)

[Publications] Ohga,N.: "Rabbit intestine contains a protein that ingibits the dissociation of GDP from and the subsequent binding fo GTP to rhoB p20,a ras p21-like GTP-binding protein." Biochem.Biophys.Res.Commun.163. 1523-1533 (1989)

[Publications] Yamamoto,K.: "Small molecular weight GTP-binding proteins and signal transduction." Clinica.Chimica.Acta.185. 347-356 (1989)

[Publications] Fukumoto,Y.: "Activation of the c-fos serum-response element by the activated c-Ha-ras protein in a manner independent of protein kinase C and cAMP-dependent protein kinase." J.Biol.Chem.265. 774-780 (1990)

[Publications] Sasaki,T.: "Purification and characterization from bovine brain cytosol of a protein that ingibits the dissociation of GDP from and the subsequent binding of GTP to smg p25A,a ras p21-like GTP-binding proein." J.Biol.Chem.265. 2333-2337 (1990)

[Publications] Hoshijima,M.: "Purification and characterization from bovine brain membranes of a GTP-binding protein with a Mr of 21,000 ADP-ribosylated by an ADP-ribosyltransferase contained in botulinum toxin type c1-Identification as the rhoA gene product." Mol.Brain Res.7. 9-16 (1990)

[Publications] Matsui,Y.: "Molecular cloning of smg p21B and identification of smg p21 purified from bovine brain and human platelets as smg p21B." Biochem.Biophys.Res.Commun.166. 1010-1016 (1990)

[Publications] Hata,Y.: "inhibition of the ras p21 GTPase-activating protein-stimulated GTPase activity of c-Ha-ras p21 by smg p21 gaving the same putative effector domain as ras p21s." J.Biol.Chem.(1990)

[Publications] Yamamoto,J.: "A GTPase-activating protein(GAP)for rhoB p20,a ras p21-like GTP-binding protein-Partial purification,characterization and subcellular distribution in rat brain." Mol.Brain Res.(1990)

[Publications] Kim,S.: "Tissue and subcellular distributions of smg p21s,ras p21-like GTP-binding proteins having the same putative effector domain as ras p21s,partly sistinct from those of c-ras p21s." Mol,Cell.Biol.(1990)

高井義美神戸大学, 医学部, 教授 (60093514)