INHIBITOR MOLECULAR BIOLOGY FOR ELUCIDATION OF CHROMATIN FUNCTION (1998)

用于阐明染色质功能的抑制剂分子生物学 (1998)

• 批准号: 09460045
• 负责人: YOSHIDA Minoru
• 金额: $ 10.37万
• 依托单位: THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09460045/
• 关键词: chromatin; histone; acetylation; nuclear export; trichostatin; leptomycin; CRM1; transcription; ヒストンアセチル化; 細胞周期; CDK

phd1^+, the first histone deacetylase gene from fission yeast Schizosaccharomyces pombe, was cloned. The gene was required for the onset of meiotic cell cycle and resistance to trichostatin A (TSA), a histone deacetylase inhibitor. RT-PCR and Western blot analyses revealed that p2lWaf1, a cyclin-dependent kinase inhibitor, was induced in mammalian cells in response to TSA.The enhanced expression was independent of p53, a major tumor suppressor protein, suggesting that p21 induction is responsible for TSA-induced cell cycle arrest. A human homolog of ORM1, the target protein of leptomycin B (LMB), was cloned. We showed that CRM1 is a receptor for the nuclear export signal (NES) of proteins, and that LMB directly binds CRM1 to inhibit its function. Using LMB, we demonstrated that Pap1, a fission yeast AP-1 like transcription factor, contained NES in the C-terminal region, which is responsible for its cytoplasmic localization.

phd1^+，第一个来自裂殖酵母裂殖酵母的组蛋白脱乙酰酶基因被克隆。该基因是减数分裂细胞周期开始和对曲古抑菌素 A (TSA)（一种组蛋白脱乙酰酶抑制剂）的抗性所必需的。 RT-PCR 和 Western blot 分析显示，p2lWaf1（一种细胞周期蛋白依赖性激酶抑制剂）在哺乳动物细胞中响应 TSA 而被诱导。表达增强与主要肿瘤抑制蛋白 p53 无关，表明 p21 诱导与 TSA 相关。 -诱导细胞周期停滞。克隆了瘦霉素 B (LMB) 靶蛋白 ORM1 的人类同源物。我们证明CRM1是蛋白质核输出信号（NES）的受体，LMB直接结合CRM1以抑制其功能。使用LMB，我们证明了Pap1（一种裂殖酵母AP-1样转录因子）在C端区域包含NES，这负责其细胞质定位。

项目成果

Kudo, N., Taoka, H., Toda, T., Yoshida, M., and Horinouchi, S.: "A novel nuclear export signal sensitive to oxidative stress in the fission yeast transcription factor Pap1." J.Biol.Chem.(in press). (1999)

Kudo, N.、Taoka, H.、Toda, T.、Yoshida, M. 和 Horinouchi, S.：“裂殖酵母转录因子 Pap1 中对氧化应激敏感的新型核输出信号。”

Yoshida et al.: "Leptomycin,an antifungal and antitumor Streptomyces metabolite,is a novel chemical probe for eukaryotic nucleo-cytoplasmic transport." Actinomycetologica. 12. 120-128 (1999)

Yoshida 等人：“Leptomycin 是一种抗真菌和抗肿瘤链霉菌代谢物，是一种用于真核细胞核质运输的新型化学探针。”

M.Fukuda et al.: "CRM1 is responsible for intracellular transport mediated by the nuclear export signal" Nature. 390. 308-311 (1997)

M.Fukuda 等人：“CRM1 负责由核输出信号介导的细胞内运输”Nature。

Ki, S.W., Kasahara, K., Kwon, H.J., Eishima, J., Takasako, K., Cooper, J.A., Yoshida, M., and Horinouchi, S.: "Identification of radicicol as an inhibitor of in vivo Ras/Raf interaction by the yeast two-hybrid screen system." J.Antibiot.51. 936-944 (1998)

Ki, S.W.、Kasahara, K.、Kwon, H.J.、Eishima, J.、Takasako, K.、Cooper, J.A.、Yoshida, M. 和 Horinouchi, S.：“鉴定根赤霉素作为体内 Ras 抑制剂/

Sano, M., Yoshida, M., Fukui, S., and Kitajima, S.: "Radicicol potentiates neurotrophin-mediated neurite outgrowth and survival of cultured sensory neurons from chick embryo." J.Neurochem.(in press). (1999)

Sano, M.、Yoshida, M.、Fukui, S. 和 Kitajima, S.：“Radicicol 增强神经营养素介导的神经突生长和鸡胚培养感觉神经元的存活。”