1993 Fiscal Year Annual Research Report

低分子量GTP結合蛋白質の作用機構と役割

Research Project

Project/Area Number	05271103
Research Institution	Gifu University
Principal Investigator	野沢義則岐阜大学, 医学部, 教授 (10021362)
Co-Investigator(Kenkyū-buntansha)	松本邦弘名古屋大学, 理学部, 教授 (70116375) 山本正幸東京大学, 大学院理系, 教授 (40114706) 石橋貞彦広島大学, 医学部, 教授 (90012616) 成宮周京都大学, 医学部, 教授 (70144350) 高井義美神戸大学, 医学部, 教授 (60093514)
Keywords	低分子量GTP結合蛋白質 / 細胞内小胞輸送 / ADPリボシル化 / リン酸化 / 酵母
Research Abstract	哺乳動物細胞における低分子量GTP結合蛋白質(LMG)の研究成果: 1)ram蛋白質の巨核球系培養細胞への発現のための発現ベクターを作成した。2)smg p25(rab3A)の標的蛋白質を精製・クローニングし、rabphilin-3Aと命名した。3)rho蛋白質の細胞内情報伝達系における役割を培養細胞を用いて検討し、三量体G蛋白質を介するチロシンリン酸化とPI3キナーゼ活性化に、rhoが関与することを明らかにした。4)未成熟ラット精巣の核内にADPリボシル化酵素およびその基質となる分子量20KDaの蛋白質を見いだし、それらの性状を検討した。5)ニワトリ末梢多形核白血球顆粒に見いだしたADPリボシル転移酵素の標的蛋白質p33がmim1(myb-induced myeloid protein-1)であることを報告し、その基質特異性について検討した。6)慢性関節リウマチ患者血清中の自己抗体によって認識され、ゴルジ装置に局在する新しい蛋白質を同定した。7)インスリン分泌において、カルシウム結合蛋白質であるカルサイクリンが促進作用を示すことを明らかにした。酵母における研究成果: 1)分裂酵母の接合フェロモンPおよびMファクターのシグナル伝達の下流で機能するSpk1がMAPキナーゼであることを示した。また、Pファクターを精製・同定した。2)rho類似蛋白質のCDC42の下流でI型フォスファターゼが機能することを明らかにした。3)小胞体-ゴルジ体間の輸送に必須なLMGであるSarlにおけるGTP水解は、小胞体からの輸送小胞の形成以降に必須であることが判明した。なお、上記以外の生物種においても、1)大腸菌における染色体の複製開始蛋白質DnaAが細胞運動に関与することが示され、また、2)繊毛虫であるテトラヒメナにおいて、二種類のLMGとしてRan/TC4がクローニングされた。

Research Products

(22 results)

All Other

All Publications (22 results)

[Publications] Nagata,K.: "Subcellular localization of a low Mr GTP-binding protein,c25KG,in resting and stimulated human platelets." Biochem.Biophys.Res.Commun.195. 1081-1088 (1993)
[Publications] Nagata,K.: "Cloning of cDNAs encoding a cell-cycle-regulatory GTP-binding low Mr (GBLM)protein,Ran/TC4,from micronucleated Tetrahymena thermophila and amicronucleated T.pyriformis." Gene. (in press). (1994)
[Publications] Shirataki,H.: "Rabphilain-3A,a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.13. 2061-2068 (1993)
[Publications] Yamaguchi,T.: "Two functionally different domains of rabphilin-3A,rab3A p25/smg p25A-binding and phospholipid- and Ca^<2+>-binding domains." J.Biol.Chem.268. 27164-27170 (1993)
[Publications] Kumagai,N.: "Lysophosphatidic acid-stimulated protein tyrosine phosphorylation in Swiss 3T3 cells." FEBS lett.329. 273-276 (1993)
[Publications] Kumagai,N.: "ADP-ribosylation of rho p21 inhibits lysophosphatidic acid-induced protein tyrosine phosphorylation and activation of MAP-kinase and focal adhesion kinase in cultured Swiss 3T3 cells" J.Biol.Chem.268. 24535-24538 (1993)
[Publications] Takeda,S.: "Purification of a GTP-binding protein localized in mitochondria." J.Biochem.114. 684-690 (1993)
[Publications] Tomura,A.: "Requirement of phosphatidylglycerol for flagellation of Escherichia coli." FEBS Lett.287. 287-290 (1993)
[Publications] Maruo,J.: "Regulation of Na^+/glucose cotransport in LLC-PK_1 cells by glucose in culture medium:involvement of Gi protein." Biochem.Mol.Biol.Int.31. 469-475 (1993)
[Publications] Yamaguchi,M.: "Cytosolic protein phosphatase may turn off activated NADPH oxidase in guinea pig neutrophils." Arch.Biochem.Biophys.306. 209-214 (1993)
[Publications] Taniguchi,M.: "Comparison of acceptor protein specificities on the formation of ADP-ribose acceptor adducts by arginine-specific ADP-ribosyltransferase from rabbit skeletal muscle sarco-plasmic reticulum with those of the enzyme from chicken peripheral polymorphonuclear cells." Biochim.Biophys.Acta. 1161. 265-271 (1993)
[Publications] Tsuchiya,M.: "Target protein for eukaryotic arginine-specific ADP-ribosyltransferase" Mol.Cell.Biochem.(in press). (1993)
[Publications] Okimoto,T.: "Brefeldin A protects ricin-induced cytotoxicity in human cancer KB cell line,but not in its resistant counterpart with altered Golgi structures." Cell Struct.Funct.18. 241-251 (1993)
[Publications] Oda,K.: "Conversion of secretory proteins into membrane proteins by fusing with a glycosylphosphatidylinositol anchor signal of alkaline phosphatase." Biochem.J.(in press). (1994)
[Publications] Okazaki,K.: "A role of calcyclin,a Ca2+-binding protein,on the Ca2+-dependent insulin release from the pancreatic β cell." J.Biol.Chem.(in press). (1994)
[Publications] Niki,I.: "Presence and possible involvement of Ca/calmodulin dependent protein kinases in insulin release from the rat pancreatic β cell." Biochem.Biophys.Res.Commun.191. 255-261 (1993)
[Publications] Gotoh,Y.: "Schizosaccharomyces pombe Spk1 is a tyrosin-phosphorylated protein functionally related to Xenopus mitogen-activated protein kinase." Mol.Cell.Biol.13. 6427-6434 (1993)
[Publications] Imai,Y.: "The fission yeast mating pheromone P-factor:its molecular structure,gene and ability to induce gene expression and G1 arrest in the mating partner." Genes & Dev.8. 328-338 (1993)
[Publications] Irie,K.: "MKK1 and MKK2,encoding Saccharomyces cerevisiae MAK kinase-kinase homologs,function in the pathway mediated by protein kinase C." Mol.Cell.Biol.13. 3076-3083 (1993)
[Publications] Doi,K.: "MSG5,a novel protein phosphatase promotes adaptation to pheromone respose in S.cerevisiae." EMBO J.13. 61-70 (1993)
[Publications] Oka,T.: "Inhibition of GTP hydrolysis by Sar1 causes accumulation of vesicles that are a functional intermediate of the ER-to Golgi transport in yeast." J.Cell Biol.(in press). (1994)
[Publications] Nishikawa,S.: "Identification of a gene required for membrane protein retention in the early secretory pathway." Proc.Natl.Acad.Sci.USA. 90. 8179-8183 (1993)

1993 Fiscal Year Annual Research Report

低分子量GTP結合蛋白質の作用機構と役割

Principal Investigator

野沢 義則 岐阜大学, 医学部, 教授 (10021362)

Research Products

[Publications] Nagata,K.: "Subcellular localization of a low Mr GTP-binding protein,c25KG,in resting and stimulated human platelets." Biochem.Biophys.Res.Commun.195. 1081-1088 (1993)

[Publications] Nagata,K.: "Cloning of cDNAs encoding a cell-cycle-regulatory GTP-binding low Mr (GBLM)protein,Ran/TC4,from micronucleated Tetrahymena thermophila and amicronucleated T.pyriformis." Gene. (in press). (1994)

[Publications] Shirataki,H.: "Rabphilain-3A,a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin." Mol.Cell.Biol.13. 2061-2068 (1993)

[Publications] Yamaguchi,T.: "Two functionally different domains of rabphilin-3A,rab3A p25/smg p25A-binding and phospholipid- and Ca^<2+>-binding domains." J.Biol.Chem.268. 27164-27170 (1993)

[Publications] Kumagai,N.: "Lysophosphatidic acid-stimulated protein tyrosine phosphorylation in Swiss 3T3 cells." FEBS lett.329. 273-276 (1993)

[Publications] Kumagai,N.: "ADP-ribosylation of rho p21 inhibits lysophosphatidic acid-induced protein tyrosine phosphorylation and activation of MAP-kinase and focal adhesion kinase in cultured Swiss 3T3 cells" J.Biol.Chem.268. 24535-24538 (1993)

[Publications] Takeda,S.: "Purification of a GTP-binding protein localized in mitochondria." J.Biochem.114. 684-690 (1993)

[Publications] Tomura,A.: "Requirement of phosphatidylglycerol for flagellation of Escherichia coli." FEBS Lett.287. 287-290 (1993)

[Publications] Maruo,J.: "Regulation of Na^+/glucose cotransport in LLC-PK_1 cells by glucose in culture medium:involvement of Gi protein." Biochem.Mol.Biol.Int.31. 469-475 (1993)

[Publications] Yamaguchi,M.: "Cytosolic protein phosphatase may turn off activated NADPH oxidase in guinea pig neutrophils." Arch.Biochem.Biophys.306. 209-214 (1993)

[Publications] Tsuchiya,M.: "Target protein for eukaryotic arginine-specific ADP-ribosyltransferase" Mol.Cell.Biochem.(in press). (1993)

[Publications] Okimoto,T.: "Brefeldin A protects ricin-induced cytotoxicity in human cancer KB cell line,but not in its resistant counterpart with altered Golgi structures." Cell Struct.Funct.18. 241-251 (1993)

[Publications] Oda,K.: "Conversion of secretory proteins into membrane proteins by fusing with a glycosylphosphatidylinositol anchor signal of alkaline phosphatase." Biochem.J.(in press). (1994)

[Publications] Okazaki,K.: "A role of calcyclin,a Ca2+-binding protein,on the Ca2+-dependent insulin release from the pancreatic β cell." J.Biol.Chem.(in press). (1994)

[Publications] Niki,I.: "Presence and possible involvement of Ca/calmodulin dependent protein kinases in insulin release from the rat pancreatic β cell." Biochem.Biophys.Res.Commun.191. 255-261 (1993)

[Publications] Gotoh,Y.: "Schizosaccharomyces pombe Spk1 is a tyrosin-phosphorylated protein functionally related to Xenopus mitogen-activated protein kinase." Mol.Cell.Biol.13. 6427-6434 (1993)

[Publications] Imai,Y.: "The fission yeast mating pheromone P-factor:its molecular structure,gene and ability to induce gene expression and G1 arrest in the mating partner." Genes & Dev.8. 328-338 (1993)

[Publications] Irie,K.: "MKK1 and MKK2,encoding Saccharomyces cerevisiae MAK kinase-kinase homologs,function in the pathway mediated by protein kinase C." Mol.Cell.Biol.13. 3076-3083 (1993)

[Publications] Doi,K.: "MSG5,a novel protein phosphatase promotes adaptation to pheromone respose in S.cerevisiae." EMBO J.13. 61-70 (1993)

[Publications] Oka,T.: "Inhibition of GTP hydrolysis by Sar1 causes accumulation of vesicles that are a functional intermediate of the ER-to Golgi transport in yeast." J.Cell Biol.(in press). (1994)

[Publications] Nishikawa,S.: "Identification of a gene required for membrane protein retention in the early secretory pathway." Proc.Natl.Acad.Sci.USA. 90. 8179-8183 (1993)

野沢義則岐阜大学, 医学部, 教授 (10021362)