2003 Fiscal Year Annual Research Report

低分子バイオプローブの開発とそれを用いた細胞内情報伝達因子のプロテオーム解析

Research Project

Project/Area Number	15310156
Research Institution	The Institute of Physical and Chemical Research
Principal Investigator	長田裕之独立行政法人理化学研究所, 抗生物質研究室, 主任研究員 (80160836)
Co-Investigator(Kenkyū-buntansha)	植木雅志独立行政法人理化学研究所, 抗生物質研究室, 研究員 (90312264) 渡邉信元独立行政法人理化学研究所, 抗生物質研究室, 先任研究員 (90221689)
Keywords	Wee1 / ユビキチン / 細胞周期 / Cdc2 / エポキシキノール
Research Abstract	プロテインキナーゼWee1は、細胞分裂期(M期)の開始・進行に中心的に働くCdc2を、M期開始の準備が整うまでリン酸化により不活性化状態に保つ役割を有する。M期開始時にヒト体細胞型Wee1(Wee1Hu/Wee1A)は、タンパク質リン酸化および分解によって不活性化されることが知られている。我々は、出芽酵母およびヒト培養細胞を用いた解析により、Wee1はユビキチンリガーゼSCF^<β-TrCP>でユビキチン化され、プロテアソーム依存に分解されることを明らかにした。Wee1AにはIκB、βカテニンなどに知られるβ-TrCPの結合コンセンサス配列(D(p)SGXX(p)S)は存在しないが、β-TrCP結合のコンセンサス配列と類似した複数の領域が結合に関与しており、領域中のセリンのリン酸化が結合に必須であることを明らかにした。さらにM期開始時に活性化されるCdc2およびPlkl(Polo like kinase 1)がこの結合に必要なセリンをリン酸化すること、これらのリン酸化酵素が実際にWee1Aとβ-TrCPの結合を促進することを確認した。低分子バイオプローブおよびRNAiを用いた実験から、Wee1Aのユビキチン化依存的な分解がM期開始に必須であることも明らかになった。また、細胞内情報伝達を制御するバイオプローブ(エポキシキノールB)をヒト血管内皮細胞に処理した際に変化する遺伝子およびタンパク質をcDNAマイクロアレイと2次元電気泳動により解析を行った。いくつかの遺伝子発現に変動が見られたので、今後、その評価を行う。

Research Products

(19 results)

All Other

All Publications (19 results)

[Publications] Watanabe, N. et al.: "M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbTrCP."Proc.Natl.Acad.Sci., USA.. (in press). (2004)
[Publications] Simizu, S. et al.: "Secretion of heparanase protein is regulated by glycosylation in human tumor cell lines."J.Biol.Chem.. 279. 2697-2703 (2004)
[Publications] Woehlecke, H. et al.: "Reversal of breast cancer resistance protein-mediated drug resistance by tryprostatin A."Int.J.Cancer.. 107. 721-728 (2003)
[Publications] Ganesh, S. et al.: "The lafora disease gene product laforin interacts with HIRIP5, a phylogenetically conserved protein containing a Nifu-like domain."Hum.Mol.Genet.. 12. 2359-2368 (2003)
[Publications] Kanoh, N. et al.: "Immobilization of natural products on glass slides by using a photoaffinity reaction and the detection of protein-small molecule interactions."Angew.Chem.Int.Ed.. 42. 5584-5587 (2003)
[Publications] Kakeya, H. et al.: "Novel non-peptide inhibitors targeting death receptor-mediated apoptosis."Bioorg.Med.Chem.Lett.. 13. 3743-3746 (2003)
[Publications] Su, S.et al.: "Synthesis and cell cycle inhibition of the peptide enamide natural products terpeptin and the aspergillamides."Tetrahedron.. 59. 8931-8946 (2003)
[Publications] Maruyama, M. et al.: "Renal abnormalities in mice Caused by insufficiency of p38 α."J.Rec.Sig.Trans.. 23. 173-183 (2003)
[Publications] Shoji, M. et al.: "Reaction modes of oxidative dimerization of epoxycyclohexenols."Tetrahedron Lett.. 44. 7205-7207 (2003)
[Publications] Palaniappan, N. et al.: "Enhancement and selective production of phoslactomycin B, a protein phosphatase iia inhibitor, through identification and engineering of the corresponding biosynthetic gene cluster."J.Biol.Chem.. 278. 35552-35557 (2003)
[Publications] Nagashima, F. et al.: "Apoptosis-inducing properties of ent-kaurene-type diterpenoids from the liverwort jungermannia truncata."Planta Medica. 69. 377-379 (2003)
[Publications] Sekiyama, Y. et al.: "Biosynthesis of phoslactomycins : cyclohexanecarboxylic acid as the starter unit."Tetrahedron. 59. 7465-7471 (2003)
[Publications] Hayashi, Y. et al.: "Asymmetric total synthesis of pseurotin A."Organic Lett.. 5. 2287-2290 (2003)
[Publications] Simizu, S. et al.: "Expression of heparanase in human tumor cell lines and human head and neck tumors."Cancer Lett.. 193. 83-89 (2003)
[Publications] Kawatani, M. et al.: "Transmembrane domain of Bcl-2 is required for inhibition of ceramide synthesis, but not cytochrome c release in the pathway of inostamysin-induced apoptosis."Exp. Cell Res.. 286. 57-66 (2003)
[Publications] Osada, H.: "Development and application of bioprobes for mammalian cell cycle analyses."Cur.Med.Chem.. 10. 727-732 (2003)
[Publications] Miyake, Y. et al.: "Epoxycyclohexenone inhibits fas-mediated apoptosis by blocking activation of pro-caspase-8 in the death-inducing signaling complex."J.Biol.Chem.. 278. 11213-11220 (2003)
[Publications] Nakazawa, J. et al.: "A novel action of terpendole E on the motor activity of mitotic kinesin Eg5."Chem. & Biol.. 10. 131-137 (2003)
[Publications] Nakamura, M. et al.: "Nordihydroguaiaretic acid, of a new family of microtubule-stabilizing agents, shows effects differentiated from paclitaxel."Biosci.Biotechnol.Biochem.. 67. 151-157 (2003)

2003 Fiscal Year Annual Research Report

低分子バイオプローブの開発とそれを用いた細胞内情報伝達因子のプロテオーム解析

Principal Investigator

長田 裕之 独立行政法人理化学研究所, 抗生物質研究室, 主任研究員 (80160836)

Research Products

[Publications] Watanabe, N. et al.: "M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbTrCP."Proc.Natl.Acad.Sci., USA.. (in press). (2004)

[Publications] Simizu, S. et al.: "Secretion of heparanase protein is regulated by glycosylation in human tumor cell lines."J.Biol.Chem.. 279. 2697-2703 (2004)

[Publications] Woehlecke, H. et al.: "Reversal of breast cancer resistance protein-mediated drug resistance by tryprostatin A."Int.J.Cancer.. 107. 721-728 (2003)

[Publications] Ganesh, S. et al.: "The lafora disease gene product laforin interacts with HIRIP5, a phylogenetically conserved protein containing a Nifu-like domain."Hum.Mol.Genet.. 12. 2359-2368 (2003)

[Publications] Kanoh, N. et al.: "Immobilization of natural products on glass slides by using a photoaffinity reaction and the detection of protein-small molecule interactions."Angew.Chem.Int.Ed.. 42. 5584-5587 (2003)

[Publications] Kakeya, H. et al.: "Novel non-peptide inhibitors targeting death receptor-mediated apoptosis."Bioorg.Med.Chem.Lett.. 13. 3743-3746 (2003)

[Publications] Su, S.et al.: "Synthesis and cell cycle inhibition of the peptide enamide natural products terpeptin and the aspergillamides."Tetrahedron.. 59. 8931-8946 (2003)

[Publications] Maruyama, M. et al.: "Renal abnormalities in mice Caused by insufficiency of p38 α."J.Rec.Sig.Trans.. 23. 173-183 (2003)

[Publications] Shoji, M. et al.: "Reaction modes of oxidative dimerization of epoxycyclohexenols."Tetrahedron Lett.. 44. 7205-7207 (2003)

[Publications] Palaniappan, N. et al.: "Enhancement and selective production of phoslactomycin B, a protein phosphatase iia inhibitor, through identification and engineering of the corresponding biosynthetic gene cluster."J.Biol.Chem.. 278. 35552-35557 (2003)

[Publications] Nagashima, F. et al.: "Apoptosis-inducing properties of ent-kaurene-type diterpenoids from the liverwort jungermannia truncata."Planta Medica. 69. 377-379 (2003)

[Publications] Sekiyama, Y. et al.: "Biosynthesis of phoslactomycins : cyclohexanecarboxylic acid as the starter unit."Tetrahedron. 59. 7465-7471 (2003)

[Publications] Hayashi, Y. et al.: "Asymmetric total synthesis of pseurotin A."Organic Lett.. 5. 2287-2290 (2003)

[Publications] Simizu, S. et al.: "Expression of heparanase in human tumor cell lines and human head and neck tumors."Cancer Lett.. 193. 83-89 (2003)

[Publications] Kawatani, M. et al.: "Transmembrane domain of Bcl-2 is required for inhibition of ceramide synthesis, but not cytochrome c release in the pathway of inostamysin-induced apoptosis."Exp. Cell Res.. 286. 57-66 (2003)

[Publications] Osada, H.: "Development and application of bioprobes for mammalian cell cycle analyses."Cur.Med.Chem.. 10. 727-732 (2003)

[Publications] Miyake, Y. et al.: "Epoxycyclohexenone inhibits fas-mediated apoptosis by blocking activation of pro-caspase-8 in the death-inducing signaling complex."J.Biol.Chem.. 278. 11213-11220 (2003)

[Publications] Nakazawa, J. et al.: "A novel action of terpendole E on the motor activity of mitotic kinesin Eg5."Chem. & Biol.. 10. 131-137 (2003)

[Publications] Nakamura, M. et al.: "Nordihydroguaiaretic acid, of a new family of microtubule-stabilizing agents, shows effects differentiated from paclitaxel."Biosci.Biotechnol.Biochem.. 67. 151-157 (2003)

長田裕之独立行政法人理化学研究所, 抗生物質研究室, 主任研究員 (80160836)