1990 Fiscal Year Annual Research Report

プロテアソ-ム(巨大蛋白分解酵素復合体)の構造解析と癌化における発現異常の解明

Research Project

Project/Area Number	02152081
Research Institution	The University of Tokushima
Principal Investigator	市原明徳島大学, 酵素センター, 教授 (40035374)
Co-Investigator(Kenkyū-buntansha)	富田優美子徳島大学, 酵素センター, 助手 (00089913) 田中啓二徳島大学, 酵素センター, 助手 (10108871) 野田千征子徳島大学, 酵素センター, 助教授 (40035506)
Keywords	プロテアソ-ム / 蛋白分解酵素 / ユビキチン / 癌胎児性蛋白 / 遺伝子破壊
Research Abstract	1.プロテアソ-ムを酵母,ラット,ヒト肝臓から精製し,そのサブユニットの数種のアミノ酸配列をcDNAで決定した。殆んどのN末端はアセチル化されていた。いづれのサブユニット構造も既知蛋白と類似性はないが,異種動物間,異種サブユニット間には類似性があり新しい蛋白族である。また數種のサブユニットにはチロシンリン酸化部位や核移行シグナルを同定した。 2.酵母の二種サブユニットcDNA飜訳領域を遺伝子破壊すると増殖しなくなる。しかし非飜訳領域の破壊は正常であった。 3.ヒト原発肝癌、腎癌,白血病,これらの樹立株ではプロテアソ-ムのサブユニットmRNAは著明に上昇し、かつ免疫染色で核に増加している。また胎生初期の組織にも高い、しかし成熱組織の増殖では変化は見られなかった。 4.プロテアソ-ムは組織抽出派からATP存在下に精製すると20Sの大きさから26Sの更に大きな粒子になる。26SにはATPase活性があり分子量4万ー10万以上の10数種の蛋白因子がプロテアソ-ムに結合している。酵素活性はユビキチン結合蛋白をATP存在下に分解することが出来る。 5.以上の結果はプロテアソ-ムが長い間知られていたエネルギ-・ユビキチン依存性蛋白分解の本態であり,しかも細胞生存,味熟状態(癌を含む)で必須の役割を演じていること,そしてその構造は新しい分子種であり動物界に広く保存されていること,これらすべての事実がその重要性を示している。

Research Products

(13 results)

All Other

All Publications (13 results)

[Publications] Akira ICHIHARA: "Regulation of protein and amino acid metabolism in cultured hepatocytes" Nutrition:Protein and Amino Acids. 49-66 (1990)
[Publications] Keiji TANAKA: "Molecular cloning of cDNA for proteasomes from rat liver:Primary strucuture of component C3 with a possible tyrosine phosphorylation site" Biochemistry. 29. 3777-3785 (1990)
[Publications] Fuminori TOkUNAGA: "The NH_2ーterminal residues of rat liver proteasome(multicatalytic proteinase complex)subunits,C2,C3 and C8,are Naーacetylated" FEBS Letter. 263. 373-375 (1990)
[Publications] Tomohiro TAMURA: "cDNA cloning and sequence of component C5 of proteasomes from rat hepatoma cells" FEBS Letter. 264. 91-94 (1990)
[Publications] Atsushi KUMATORI: "cDNA cloning and sequencing of component C9 of proteasomes from rat hepatoma cells" FEBS Letter. 264. 279-282 (1990)
[Publications] Keiji TANAKA: "Proteasomes(multicatalytic proteinase complex)in eukaryotic cells" Cell Struct.Funct,. 15. 127-132 (1990)
[Publications] Keiji TANAKA: "cDNA cloning and sequencing of component C8 of proteasomes from rat hepatoma cells" Biochem.Biophys.Res.Commun.171. 676-683 (1990)
[Publications] Tsutomu FUJIWARA: "Proteasomes are essential for yeast proliferation cDNA cloning and gene disruption of two major subunits" J.Biol.Chem.265. 16604-16613 (1990)
[Publications] Atsushi KUMATORI: "Abnormally high expression of proteasomes in human leukemic cells" Proc.Natl.Acad.Sci.USA. 87. 7071-7078 (1990)
[Publications] Keiji TANAKA: "Possible mechanism of nuclear translocation of proteasomes" FEBS Letter. 271. 41-46 (1990)
[Publications] Hiroーomi KANAYAMA: "Expression of ubiquitin genes in rat liver during development and carcinogenesis" Cancer Res.(1991)
[Publications] Tsutomu ANDO: "Developmental and cancerous changes of proteasome expression in liver" Cancer Res.(1991)
[Publications] Etsuko ORINO: "ATPーdependent reversible association of proteasomes with multiple protein factors to form 26S complexes that degrade ubiquitinated proteins in human HLー60 cells" Proc.Natl.Acad.Sci.USA. (1991)

1990 Fiscal Year Annual Research Report

プロテアソ-ム(巨大蛋白分解酵素復合体)の構造解析と癌化における発現異常の解明

Principal Investigator

市原 明 徳島大学, 酵素センター, 教授 (40035374)

Research Products

[Publications] Akira ICHIHARA: "Regulation of protein and amino acid metabolism in cultured hepatocytes" Nutrition:Protein and Amino Acids. 49-66 (1990)

[Publications] Keiji TANAKA: "Molecular cloning of cDNA for proteasomes from rat liver:Primary strucuture of component C3 with a possible tyrosine phosphorylation site" Biochemistry. 29. 3777-3785 (1990)

[Publications] Fuminori TOkUNAGA: "The NH_2ーterminal residues of rat liver proteasome(multicatalytic proteinase complex)subunits,C2,C3 and C8,are Naーacetylated" FEBS Letter. 263. 373-375 (1990)

[Publications] Tomohiro TAMURA: "cDNA cloning and sequence of component C5 of proteasomes from rat hepatoma cells" FEBS Letter. 264. 91-94 (1990)

[Publications] Atsushi KUMATORI: "cDNA cloning and sequencing of component C9 of proteasomes from rat hepatoma cells" FEBS Letter. 264. 279-282 (1990)

[Publications] Keiji TANAKA: "Proteasomes(multicatalytic proteinase complex)in eukaryotic cells" Cell Struct.Funct,. 15. 127-132 (1990)

[Publications] Keiji TANAKA: "cDNA cloning and sequencing of component C8 of proteasomes from rat hepatoma cells" Biochem.Biophys.Res.Commun.171. 676-683 (1990)

[Publications] Tsutomu FUJIWARA: "Proteasomes are essential for yeast proliferation cDNA cloning and gene disruption of two major subunits" J.Biol.Chem.265. 16604-16613 (1990)

[Publications] Atsushi KUMATORI: "Abnormally high expression of proteasomes in human leukemic cells" Proc.Natl.Acad.Sci.USA. 87. 7071-7078 (1990)

[Publications] Keiji TANAKA: "Possible mechanism of nuclear translocation of proteasomes" FEBS Letter. 271. 41-46 (1990)

[Publications] Hiroーomi KANAYAMA: "Expression of ubiquitin genes in rat liver during development and carcinogenesis" Cancer Res.(1991)

[Publications] Tsutomu ANDO: "Developmental and cancerous changes of proteasome expression in liver" Cancer Res.(1991)

[Publications] Etsuko ORINO: "ATPーdependent reversible association of proteasomes with multiple protein factors to form 26S complexes that degrade ubiquitinated proteins in human HLー60 cells" Proc.Natl.Acad.Sci.USA. (1991)

市原明徳島大学, 酵素センター, 教授 (40035374)