1990 Fiscal Year Annual Research Report

細胞増殖シグナルの伝達・発現とその制御

Research Project

Project/Area Number	02151080
Research Institution	Tokyo Metropolitan Organization for Medical Research
Principal Investigator	鈴木紘一東京都臨床医学総合研究所, 遺伝情報研究部, 部長 (80011948)
Co-Investigator(Kenkyū-buntansha)	伊庭英夫東京大学, 医科学研究所, 助教授 (60111449) 山本正幸東京大学, 理学部, 教授 (40114706) 蛯名洋介徳島大学, 酵素科学研究センター, 教授 (00112227) 宇野功東京大学, 応用微生物学研究所, 教授 (60114401) 野沢義則岐阜大学, 医学部, 教授 (10021362)
Keywords	インスリンリセプタ- / 情報伝達 / プロテインキナ-ゼC
Research Abstract	1.細胞膜近辺の現象の解析ーーインスリンリセプタ-の発現系を使いPI3キナ-ゼの85Kサブユニットがリン酸化されることが判明した。また、カエルの卵の系でM期に活性化されるキナ-ゼがMAPキナ-ゼであることを始めて明らかにし,そのcDNAのクロ-ニングを完成した。このキナ-ゼはG_0/G_1期にも特異的に活性化され,微小管と情報伝達の関係を明らかにする手がかりとしてさらに研究を進めている。情報伝達に伴い活性化されるPKCの新しい分子種をさらに数種類同定し,結局,PKCは10種類を越えるファミリ-を形成していることが明らかになった。また,これらの新しいPKC,nPKCがGH4C1細胞では生理的な刺激に反応し,情報を伝達することが,甲状腺刺激ホルモン放出ホルモンを用いた実験で明らかになった。また,PKC分子種間ではPKCγのみがホルボ-ルエステル刺激に反応してTREーCATの発現が誘導できず,PKC分子に機能分担があることが判明した。 2.核内でおこる現象の解明ーーFos,Junおよびその関連のFra1,Fra2について解析した。Jun/Fosだけでなく,Fra2/JunなどもAPー1配列に特異的に結合することが明らかになった。また,カルパスタチンのcDNAを細胞内で発現させると,JunによるTREーCATの転写活性の著しい増加が観察され,Junなどの転手因子の活性がプロテア-ゼによって制御されていることが示唆された。 3.酵母系をもとにした情報の流れの解析ーーPKCの阻害剤スタウロスポリンに対する耐性出芽酵母変異株を単離し,そのうちの1つを解析し,PKCの類似体であるものが同定された。この変異株は2nの状態で増殖を停止していた。また,分裂酵母でもアデニル酸シクラ-ゼを同定し構造を決定した。このものの遺伝子を破壊しても生育は可能であったが,有性生殖に対しては脱抑制されていた。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Akita,Y.: "Expression and properties of two distinct classes of the phorbol ester receptor family,four conventional protein kinase C types and a novel protein Kinase C." J.Biol.Chem.265. 354-362 (1990)
[Publications] Ohno,S.: "A point mutation at the putative ATPーbinding site of protein kinase C alpha abolishes the kinase activity and renders it down regulation insensitive." J.Biol.Chem.265. 6296-6300 (1990)
[Publications] Akita,Y.: "Possible role of Ca^<2+>ーindependent protein kinase C isozyme,nPKC epsilon,in thyrotropin releasing hormoneーstimulated signal transduction:differential down regulation of nPKCe in GH4C1 cells." Biochem.Biophys.Res.Commun.172. 184-189 (1990)
[Publications] Osada,S.: "A phorbol ester receptor/kinase C eta,a new member of the protein kinase C family predominantly expressed in lung and skin." J.Biol.Chem.265. 22434-22440 (1990)
[Publications] Homma,Y.: "Isolation and characterization of a gamma type phosphoinositideーspecific phospholipase C (PLC gamma 2)." Biochem.J.269. 13-18 (1990)
[Publications] Uno,I.: "The functional domain of adenylate cyclase associated with entry into myosis of <Saccharomyces cerevisiae>___ー." J.Bacteriol.172. 102-109 (1990)
[Publications] Nishina,H.: "Isolation and characterization of fraー2:an additional member of fos gene family." Proc.Natl.Acad.Sci.87. 3619-3632 (1990)
[Publications] Gotoh,Y.: "Microtubule associated protein (MAP) kinase activated by nerve growth factor and epidermal growth factor in PC12 cells." Eur.J.Biochem.193. 661-669 (1990)
[Publications] Gotoh,Y.: "Okadaic acid activates microtubule associated protein kinase in quiescent fibloblastic cells." Eur.J.Biochem.193. 671-674 (1990)
[Publications] Gotoh,Y.: "In vitro effects of microtubule dymanics of purified Xenopus M phaseーactivated MAP kinase" Nature. 349. 251-254 (1991)
[Publications] Maeda,T.: "Adenylate cyclase is dispensable for vegetative cell growth in the fission yeast <Schizosaccharomyces pombe>___ー" Proc.Natl.Acad.Sci.87. 7814-7818 (1990)
[Publications] Nagata,K.: "The ram:a novel low molecular weight GTPーbinding protein cDNA from a rat megakaryocyte library." FEBS lett.275. 29-32 (1990)
[Publications] Shmma,Y.: "Isolation and characterization of neomycinーsensitive mutant in Saccharomyces cerevisiae." J.Gen.Microbiol.136. 1753-1761 (1990)
[Publications] Murata,A.: "The adenylate cyclase/protein kinase cascade regulates entry into meiosis in <Saccharomyces cerevisiae>___ー through the gene IME1" EMBO J.9. 3225-3232 (1990)
[Publications] Shimada,F.: "Insulin resistant disbetes associated with partial deletion of insulin deletion gene." Lancet. 335. 1791-1181 (1990)
[Publications] Kitayama,H.: "Genetic analysis of the Krevー1 gene:potentiation of its tumor suppressor activity by specific poin mutations." Proc.Natl.Acad.Sci.87. 4284-4288 (1990)
[Publications] Kitayama,H.: "A domain responsible for the transformation supressor activity in Krevー1 protein" Cancer Res.81. 445-448 (1990)
[Publications] Zang,K.: "Identification of small clusters of divergent amino acids that mediate the opposing effects of ras and Krevー1" Science. 249. 162-165 (1990)

1990 Fiscal Year Annual Research Report

細胞増殖シグナルの伝達・発現とその制御

Principal Investigator

鈴木 紘一 東京都臨床医学総合研究所, 遺伝情報研究部, 部長 (80011948)

Research Products

[Publications] Akita,Y.: "Expression and properties of two distinct classes of the phorbol ester receptor family,four conventional protein kinase C types and a novel protein Kinase C." J.Biol.Chem.265. 354-362 (1990)

[Publications] Ohno,S.: "A point mutation at the putative ATPーbinding site of protein kinase C alpha abolishes the kinase activity and renders it down regulation insensitive." J.Biol.Chem.265. 6296-6300 (1990)

[Publications] Akita,Y.: "Possible role of Ca^<2+>ーindependent protein kinase C isozyme,nPKC epsilon,in thyrotropin releasing hormoneーstimulated signal transduction:differential down regulation of nPKCe in GH4C1 cells." Biochem.Biophys.Res.Commun.172. 184-189 (1990)

[Publications] Osada,S.: "A phorbol ester receptor/kinase C eta,a new member of the protein kinase C family predominantly expressed in lung and skin." J.Biol.Chem.265. 22434-22440 (1990)

[Publications] Homma,Y.: "Isolation and characterization of a gamma type phosphoinositideーspecific phospholipase C (PLC gamma 2)." Biochem.J.269. 13-18 (1990)

[Publications] Uno,I.: "The functional domain of adenylate cyclase associated with entry into myosis of <Saccharomyces cerevisiae>___ー." J.Bacteriol.172. 102-109 (1990)

[Publications] Nishina,H.: "Isolation and characterization of fraー2:an additional member of fos gene family." Proc.Natl.Acad.Sci.87. 3619-3632 (1990)

[Publications] Gotoh,Y.: "Microtubule associated protein (MAP) kinase activated by nerve growth factor and epidermal growth factor in PC12 cells." Eur.J.Biochem.193. 661-669 (1990)

[Publications] Gotoh,Y.: "Okadaic acid activates microtubule associated protein kinase in quiescent fibloblastic cells." Eur.J.Biochem.193. 671-674 (1990)

[Publications] Gotoh,Y.: "In vitro effects of microtubule dymanics of purified Xenopus M phaseーactivated MAP kinase" Nature. 349. 251-254 (1991)

[Publications] Maeda,T.: "Adenylate cyclase is dispensable for vegetative cell growth in the fission yeast <Schizosaccharomyces pombe>___ー" Proc.Natl.Acad.Sci.87. 7814-7818 (1990)

[Publications] Nagata,K.: "The ram:a novel low molecular weight GTPーbinding protein cDNA from a rat megakaryocyte library." FEBS lett.275. 29-32 (1990)

[Publications] Shmma,Y.: "Isolation and characterization of neomycinーsensitive mutant in Saccharomyces cerevisiae." J.Gen.Microbiol.136. 1753-1761 (1990)

[Publications] Murata,A.: "The adenylate cyclase/protein kinase cascade regulates entry into meiosis in <Saccharomyces cerevisiae>___ー through the gene IME1" EMBO J.9. 3225-3232 (1990)

[Publications] Shimada,F.: "Insulin resistant disbetes associated with partial deletion of insulin deletion gene." Lancet. 335. 1791-1181 (1990)

[Publications] Kitayama,H.: "Genetic analysis of the Krevー1 gene:potentiation of its tumor suppressor activity by specific poin mutations." Proc.Natl.Acad.Sci.87. 4284-4288 (1990)

[Publications] Kitayama,H.: "A domain responsible for the transformation supressor activity in Krevー1 protein" Cancer Res.81. 445-448 (1990)

[Publications] Zang,K.: "Identification of small clusters of divergent amino acids that mediate the opposing effects of ras and Krevー1" Science. 249. 162-165 (1990)

鈴木紘一東京都臨床医学総合研究所, 遺伝情報研究部, 部長 (80011948)