2000 Fiscal Year Annual Research Report

ユビキチンリガーゼを利用した癌遺伝子産物の分解促進による癌治療

Research Project

Project/Area Number	12480192
Research Institution	Hamamatsu University School of Medicine
Principal Investigator	北川雅敏浜松医科大学, 医学部, 教授 (50294971)
Co-Investigator(Kenkyū-buntansha)	内田千晴浜松医科大学, 医学部, 助手 (60223567) 小田敏明浜松医科大学, 医学部, 助教授 (90126805)
Keywords	p27^<kip1> / ユビキチン-プロテアソーム / ユビキチンリガーゼ / 細胞周期 / リン酸化
Research Abstract	1.ユビキチン結合酵素E2-Cの制御機構の解析細胞癌化にも関連し、分裂期に必須なE2-CはユビキチンリガーゼAPC/サイクロソームによって量的制御を受けていることを発見した(Yamanaka et al.Mol.Biol.Cell.,2000)。 2.p27^<Kip1>の分解機構の解析 in vitroおよびin vivoのp27^<Kip1>の分解系を確立した。この解析によりp27^<Kip1>がSkp2をユビキチンリガーゼとしたユビキチン-プロテアソーム系と限定分解の二つの経路で分解されることを見出した。マイクロシークエンスによりCDK結合部位とサイクリン結合部位の二つの限定分解部位の決定に成功した。限定分解酵素の精製方法もほぼ確立した。また、p27^<Kip1>のリン酸化部位の解析を行い、新規部位Ser10を見出した。Ser10のリン酸化あるいはそれをミミックしたGluへの変換によりp27^<Kip1>は分解耐性になる(Ishida et al.J Biol Chem,2000)。 3.p27^<Kip1>の機能と分解の個体レベルでの解析 p27^<Kip1>のユビキチンリガーゼSkp2のノックアウトマウスを作成して解析した。マウスは正常に発生するが、矮小化、肝臓、腎臓、肺胞細胞の多倍数体化、中心体の過剰複製、p27^<Kip1>およびサイクリンEの蓄積が見られた(Nakayama et al.EMBO J,2000)。p27^<Kip1>の分解は低下しているSkp2ノックアウトマウス由来の繊維芽細胞に、アデノウイルスに組み込んだSkp2を感染させることでp27^<Kip1>の分解が誘導された。このことより、Skp2ユビキチンリガーゼを阻害することでp27^<Kip1>の分解を抑制することができることが示唆された。また、p27のトランスジェニックマウスの解析結果からT細胞の分化にp27^<Kip1>の発現低下が重要であることが証明された(Tsukiyama et al.J immunol,2001)。

Research Products
(13 results)

All Other

All Publications (13 results)

[Publications] Tsukiyama,T. et al.: "Down-regulation of p27^<Kip1> expression is required for development and function of T cells."Journal of Immunology. 116. 304-312 (2001)
[Publications] Yamanaka,A. et al.: "Cell-cycle dependent expression of mammalian E2-C regulated by the anaphase promoting complex/cyclosome."Molecular Biology of the Cell. 11. 2821-2831 (2000)
[Publications] Ishida,N. et al.: "Phosphorylation at Ser10 a major phosphorylation site of p27^<Kip1>, increases its protein stability"Journal of Biological Chemistry. 275. 25146-25154 (2000)
[Publications] Ishimi,Y. et al.: "Inhibition of Mcm4, 6, 7 helicase activity by phosphorylation with cyclin A-Cdk2"Journal of Biological Chemistry. 275. 16235-16241 (2000)
[Publications] Nakayama,K. et al.: "Targeted disruption of Skp2 result in accumulation of Cyclin E and p27^<Kip1>, polyploide, and centrosome overdeplication"EMBO Journal. 19. 2069-2081 (2000)
[Publications] Dobashi,Y. et al.: "Simultaneous suppression of Cdc2/Cdk2 activity is necessary and sufficient for neuronal differentiation of PC12."Journal of Biological Chemistry. 275. 12572-12580 (2000)
[Publications] Tanaka,T. et al.: "Geranylgeranyl-pyrophosphate(GGPP), an isoprenoid of mevalonate cascade, is a critical compound for rat primary cultured cortical neurons to protect the cell death induced by HMG-CoA reductase inhibition."Journal of Neuroscience. 20. 2852-2859 (2000)
[Publications] Shimohata,T. et al.: "Expanded polyglutamine stretches associated with CAG repeat diseases interact with hTAFll130, interfering with CREB-dependent transcription"Nature Genetics. 26. 29-36 (2001)
[Publications] Xue,H-H. et al.: "Store depletion by caffeine/ryanodine activates capacitative Ca2+ entry in nonexcitable A549 cells."Journal of Biochemistry. 128. 329-336 (2000)
[Publications] Zhao,D-M. et al.: "Effect of erythromycin on ATP-induced intracellular calcium response in A549 cells"Journal of Physiology. 278. L726-L736 (2000)
[Publications] Oda,T. et al.: "Mitochondrial targeting signal-induced conformational change and repression of the peroxisomal targeting signal of the precursor for rat liver serine : pyruvate/alanine : glyoxylate aminotransferase."Journal of Biochemistry. 127. 665-671 (2000)
[Publications] Yokota,S. et al.: "Aggregate formation and degradation of overexpressed wild-type and mutant urate oxidase proteins.quality control af organelle-destined proteins by the endoplasmic reticulum"Histochemstry and Cell Biology. 114. 433-446 (2000)
[Publications] 北川雅敏(分担執筆): "わかる細胞周期と癌"田矢洋一編羊土社. 118 (2000)

2000 Fiscal Year Annual Research Report

ユビキチンリガーゼを利用した癌遺伝子産物の分解促進による癌治療

Principal Investigator

北川 雅敏 浜松医科大学, 医学部, 教授 (50294971)

Research Products

[Publications] Tsukiyama,T. et al.: "Down-regulation of p27^<Kip1> expression is required for development and function of T cells."Journal of Immunology. 116. 304-312 (2001)

[Publications] Yamanaka,A. et al.: "Cell-cycle dependent expression of mammalian E2-C regulated by the anaphase promoting complex/cyclosome."Molecular Biology of the Cell. 11. 2821-2831 (2000)

[Publications] Ishida,N. et al.: "Phosphorylation at Ser10 a major phosphorylation site of p27^<Kip1>, increases its protein stability"Journal of Biological Chemistry. 275. 25146-25154 (2000)

[Publications] Ishimi,Y. et al.: "Inhibition of Mcm4, 6, 7 helicase activity by phosphorylation with cyclin A-Cdk2"Journal of Biological Chemistry. 275. 16235-16241 (2000)

[Publications] Nakayama,K. et al.: "Targeted disruption of Skp2 result in accumulation of Cyclin E and p27^<Kip1>, polyploide, and centrosome overdeplication"EMBO Journal. 19. 2069-2081 (2000)

[Publications] Dobashi,Y. et al.: "Simultaneous suppression of Cdc2/Cdk2 activity is necessary and sufficient for neuronal differentiation of PC12."Journal of Biological Chemistry. 275. 12572-12580 (2000)

[Publications] Tanaka,T. et al.: "Geranylgeranyl-pyrophosphate(GGPP), an isoprenoid of mevalonate cascade, is a critical compound for rat primary cultured cortical neurons to protect the cell death induced by HMG-CoA reductase inhibition."Journal of Neuroscience. 20. 2852-2859 (2000)

[Publications] Shimohata,T. et al.: "Expanded polyglutamine stretches associated with CAG repeat diseases interact with hTAFll130, interfering with CREB-dependent transcription"Nature Genetics. 26. 29-36 (2001)

[Publications] Xue,H-H. et al.: "Store depletion by caffeine/ryanodine activates capacitative Ca2+ entry in nonexcitable A549 cells."Journal of Biochemistry. 128. 329-336 (2000)

[Publications] Zhao,D-M. et al.: "Effect of erythromycin on ATP-induced intracellular calcium response in A549 cells"Journal of Physiology. 278. L726-L736 (2000)

[Publications] Oda,T. et al.: "Mitochondrial targeting signal-induced conformational change and repression of the peroxisomal targeting signal of the precursor for rat liver serine : pyruvate/alanine : glyoxylate aminotransferase."Journal of Biochemistry. 127. 665-671 (2000)

[Publications] Yokota,S. et al.: "Aggregate formation and degradation of overexpressed wild-type and mutant urate oxidase proteins.quality control af organelle-destined proteins by the endoplasmic reticulum"Histochemstry and Cell Biology. 114. 433-446 (2000)

[Publications] 北川雅敏(分担執筆): "わかる細胞周期と癌"田矢洋一編 羊土社. 118 (2000)

北川雅敏浜松医科大学, 医学部, 教授 (50294971)

[Publications] 北川雅敏(分担執筆): "わかる細胞周期と癌"田矢洋一編羊土社. 118 (2000)