1989 年度実績報告書

細胞増殖シグナルの伝達とその制御

研究課題

研究課題/領域番号	01010083
研究機関	(財)東京都臨床医学総合研究所
研究代表者	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)
研究分担者	野田亮理化学研究所, ライフサイエンス筑波研究センター, 研究員 (30146708) 蛯名洋介徳島大学, 酵素科学センター, 教授 (00112227) 山本正幸東京大学, 理学部, 教授 (40114706) 宇野功東京大学, 応用微生物研究所, 教授 (60114401) 野沢義則岐阜大学, 医学部, 教授 (10021362)
キーワード	細胞増殖 / シグナル伝達 / リセプター / プロテインキナーゼC
研究概要	細胞表面に入力したシグナルが遺伝子の発現を調節するまでの過程を種種のステップに分け、各段階に関与する酵素の構造と機能をまず解明し、酵母系の知見を対比参考にしつつ各段階を再構成し、細胞のシグナル伝達機構を解明する2Cを目標に研究を展開し、今年度はその初年度である。リセプターの次のステップの解明のため、インスリンリセプターとグルコーストランスポーターを結ぶ因子の分離を試みた。両者を発現しているがインスリンに反応しない細胞を分離し、これを用いて検索をしている。リセプターの下流に位置するGタンパク質はNIH3T3を正常に復帰させるKrev1と相同性が高く、Gタンパク質とKrev1の機能を同時に検討した。GTP結合部位、エフェクター結合部位、膜結合部位に変異を導入するとKrev1の活性は著しく低下したが、Gly-12をValへ、Gln-63をGluに変換したものは逆に活性が上昇した。ホスホリパーゼCについては、新しい分子種(IV,γ2)を同定し、構造を決定すると共に、I〜IVのホスホリパーゼCについて活性発現に必須な2つの部位と、活性の修飾に必要なガン遺伝子に類似の部位を同定した。ホスホリパーゼCによって生じるジアシルグリセロールの産生は一相性であるが、細胞では2相性となりホスフアチジルコリンに由来するジアシルグリセロールの存在が証明された。Cキナーゼの分子種間の差を遺伝子の発現を目印に調べ、γがTRE-CATの発現を誘導しないことを発見した。Cキナーゼ分子種間で見出された最初の差であり、詳細な検討を進めている。Cキナーゼの下流で遺伝子発現を制御するFOSはJUNと複合体を形成して作用することを発見し、またFOSと類似性が高く免疫で交叉を示すタンパク質FRA2を見出した。酵母の系でもCAMPを介する系、形態異常を示す系でシグナルの伝達と制御を調べている。

研究成果
(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] A.Hata-Tanaka: "Spin-labeling proton NMR study on aromatic amino acid residues in the guanine nucleotide binding site of human c-Ha-ras(1-171)protein" Biochemistry. 28. 9550-9556 (1989)
[文献書誌] T.Nakamura: "Molecular cloning and expression of human hepatocyte growth factor" Nature. 342. 440-443 (1989)
[文献書誌] T.Kinoshita: "Marked increase of HGF mRNA in non-prenchymal liver cells of rats treated with hepatotoxin" Biochem.biophys.Res.Commun.165. 1229-1234 (1989)
[文献書誌] T.Asano: "Rabbit brain glucose transporter resp=onds to insulin when expressed in insulin-insensitive chinese hamster ovary cells" J.Biol.Chem.,. 264. 3416-3420 (1989)
[文献書誌] A.Hata: "Direct evidence that the kinase activity of protein kinase C is involved in transcriptional activation through a TPA-responsive" FEBS Letters. 252. 144-146 (1989)
[文献書誌] Y.Konno: "Enzymatic properties of a novel phorbol ester receptor/kinase protein kinase c,nPKC" J.Biochem.106. 673-678 (1989)
[文献書誌] Y.Homma: "Tissue and cell specific expression of mRNAs for four types of inositol-phospholipid specific phospholipase C" Biochem.Biophys.Res.Commun.164. 406-412 (1989)
[文献書誌] Y.Emori: "A second type of rat phosphoinositide specific phospholipase C containing a src-related sequence not essential for phosphoinositide hydrolyzing activity" J.Biol.Chem.264. 21885-21890 (1989)
[文献書誌] S.Tominaga: "Interferon alters the pattern of secreted proteins from Ehrlich ascites tumor cells" Biochem.J.261. 57-61 (1989)
[文献書誌] S.Tomimaga: "A putative protein of a growth specific cDNA from BALB/c-3T3 cells is highly similar to the extracellular portion of mouse interleukin 1 receptor" FEBS Letters. 258. 301-304 (1989)
[文献書誌] M.Noda: "Genes with potential for suppressing malignant phenotypes of kirsten sarcoma virus-transformed NIH-3T3 cells" UCLA Symposium on Molecular and Cellular Biology. 103. 223-229 (1989)
[文献書誌] Y.Shinohara: "Initiation of DNA synthesis by microtubule disruption in quiescent rat 3Y1 cells" Eur.J.Biochem.183. 275-280 (1989)
[文献書誌] Y.Miyata: "Protein kinase C dependent and independent pathways in the growth factor induced cytoskeletal recognition" J.Biol.chem.264. 15565-15568 (1989)
[文献書誌] M.Hoshi: "Characterization of a mitogen-activated,Ca^<2+>-sensitive microtubule-associated protein kinase" Eur.J.Biochem.184. 477-486 (1989)
[文献書誌] T.Hara: "Isolation and characterization of mouse FM3A cell mutants which are devoid of Newcastle virus receptors" J.Virol.63. 182-188 (1989)
[文献書誌] T.Murakami: "Tissue and developmental stage specific expression of the rat ornithine carbamoyltransferase gene in transgenic mice" Developmental Genetics. 10. 393-401 (1989)
[文献書誌] T.Fu: "Bradykinin-induced generation of inositol 1,4,5-triphosphate in fibloblasts and neuroblastoma cells:Effect of pertussis toxin,extracellular calcium and down regulation of protein kinase C" Biochem.Biophys.Res.Commun.157. 1429-1435 (1989)
[文献書誌] Y.Yada: "Signal transduction in the onset of terminal kerationcyte differentiation induced by 1 alpha,25-dihydroxyvitamine D3:Role of protein kinase C translocation" Biochem.Biophys.Res.Commun.163. 1517-1522 (1989)
[文献書誌] H.Kato: "Activation of phosphtidyl-inositol kinase and phosphatidyl-inositol 4-phosphate kinase by cAMP in Saccharomyces cerevisiae" J.Biol.Chem.264. 3116-3121 (1989)
[文献書誌] H.Mitsuzawa: "Isolation and characterization of temperature sensitive mutants in the Ras2 and Cyr1 genes of Sacharomyces cerevisiae" Genetics. 123. 739-748 (1989)
[文献書誌] T.Takeda: "Substitution at position 116 of schizosaccharomyces pombe calmodulin decreases its stability under nitrogen starvation and results in a sporulation-deficient phenotype" Proc.Natl.Acad.Sci.,USA. 86. 5617-5622 (1989)
[文献書誌] Y.Yoshida: "Identification of a small region of v-fos gene product that is sufficient for transforming potential and growth stimulating activity" Oncogene. (1990)
[文献書誌] T.Sekiguchi: "Insulin receptor and altered glucose transport in a monesin-resistant mutant of chinese hamster ovary cell" J.Cell Physiol.139. 229-236 (1989)
[文献書誌] M.Taira: "Human diabetes associated with a deletion of the tyrosine kinase domain of the insulin receptor" Science. 245. 63-66 (1989)

1989 年度 実績報告書

細胞増殖シグナルの伝達とその制御

研究代表者

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)

研究成果

[文献書誌] A.Hata-Tanaka: "Spin-labeling proton NMR study on aromatic amino acid residues in the guanine nucleotide binding site of human c-Ha-ras(1-171)protein" Biochemistry. 28. 9550-9556 (1989)

[文献書誌] T.Nakamura: "Molecular cloning and expression of human hepatocyte growth factor" Nature. 342. 440-443 (1989)

[文献書誌] T.Kinoshita: "Marked increase of HGF mRNA in non-prenchymal liver cells of rats treated with hepatotoxin" Biochem.biophys.Res.Commun.165. 1229-1234 (1989)

[文献書誌] T.Asano: "Rabbit brain glucose transporter resp=onds to insulin when expressed in insulin-insensitive chinese hamster ovary cells" J.Biol.Chem.,. 264. 3416-3420 (1989)

[文献書誌] A.Hata: "Direct evidence that the kinase activity of protein kinase C is involved in transcriptional activation through a TPA-responsive" FEBS Letters. 252. 144-146 (1989)

[文献書誌] Y.Konno: "Enzymatic properties of a novel phorbol ester receptor/kinase protein kinase c,nPKC" J.Biochem.106. 673-678 (1989)

[文献書誌] Y.Homma: "Tissue and cell specific expression of mRNAs for four types of inositol-phospholipid specific phospholipase C" Biochem.Biophys.Res.Commun.164. 406-412 (1989)

[文献書誌] Y.Emori: "A second type of rat phosphoinositide specific phospholipase C containing a src-related sequence not essential for phosphoinositide hydrolyzing activity" J.Biol.Chem.264. 21885-21890 (1989)

[文献書誌] S.Tominaga: "Interferon alters the pattern of secreted proteins from Ehrlich ascites tumor cells" Biochem.J.261. 57-61 (1989)

[文献書誌] S.Tomimaga: "A putative protein of a growth specific cDNA from BALB/c-3T3 cells is highly similar to the extracellular portion of mouse interleukin 1 receptor" FEBS Letters. 258. 301-304 (1989)

[文献書誌] M.Noda: "Genes with potential for suppressing malignant phenotypes of kirsten sarcoma virus-transformed NIH-3T3 cells" UCLA Symposium on Molecular and Cellular Biology. 103. 223-229 (1989)

[文献書誌] Y.Shinohara: "Initiation of DNA synthesis by microtubule disruption in quiescent rat 3Y1 cells" Eur.J.Biochem.183. 275-280 (1989)

[文献書誌] Y.Miyata: "Protein kinase C dependent and independent pathways in the growth factor induced cytoskeletal recognition" J.Biol.chem.264. 15565-15568 (1989)

[文献書誌] M.Hoshi: "Characterization of a mitogen-activated,Ca^<2+>-sensitive microtubule-associated protein kinase" Eur.J.Biochem.184. 477-486 (1989)

[文献書誌] T.Hara: "Isolation and characterization of mouse FM3A cell mutants which are devoid of Newcastle virus receptors" J.Virol.63. 182-188 (1989)

[文献書誌] T.Murakami: "Tissue and developmental stage specific expression of the rat ornithine carbamoyltransferase gene in transgenic mice" Developmental Genetics. 10. 393-401 (1989)

[文献書誌] T.Fu: "Bradykinin-induced generation of inositol 1,4,5-triphosphate in fibloblasts and neuroblastoma cells:Effect of pertussis toxin,extracellular calcium and down regulation of protein kinase C" Biochem.Biophys.Res.Commun.157. 1429-1435 (1989)

[文献書誌] Y.Yada: "Signal transduction in the onset of terminal kerationcyte differentiation induced by 1 alpha,25-dihydroxyvitamine D3:Role of protein kinase C translocation" Biochem.Biophys.Res.Commun.163. 1517-1522 (1989)

[文献書誌] H.Kato: "Activation of phosphtidyl-inositol kinase and phosphatidyl-inositol 4-phosphate kinase by cAMP in Saccharomyces cerevisiae" J.Biol.Chem.264. 3116-3121 (1989)

[文献書誌] H.Mitsuzawa: "Isolation and characterization of temperature sensitive mutants in the Ras2 and Cyr1 genes of Sacharomyces cerevisiae" Genetics. 123. 739-748 (1989)

[文献書誌] T.Takeda: "Substitution at position 116 of schizosaccharomyces pombe calmodulin decreases its stability under nitrogen starvation and results in a sporulation-deficient phenotype" Proc.Natl.Acad.Sci.,USA. 86. 5617-5622 (1989)

[文献書誌] Y.Yoshida: "Identification of a small region of v-fos gene product that is sufficient for transforming potential and growth stimulating activity" Oncogene. (1990)

[文献書誌] T.Sekiguchi: "Insulin receptor and altered glucose transport in a monesin-resistant mutant of chinese hamster ovary cell" J.Cell Physiol.139. 229-236 (1989)

[文献書誌] M.Taira: "Human diabetes associated with a deletion of the tyrosine kinase domain of the insulin receptor" Science. 245. 63-66 (1989)

1989 年度実績報告書

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)