Development of methods for using fission yeast as a test tube for analyzing highly complex biological systems.

开发使用裂殖酵母作为试管来分析高度复杂的生物系统的方法。

• 批准号: 07557196
• 负责人: OKAYAMA Hiroto
• 金额: $ 2.75万
• 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 (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557196/
• 关键词: differentiation; cell proliferation; fission yeast; stell; switching; RNA binding protein; stell; RNA 結合蛋白; 機能相補; cDNAバンク; 選択マーカー; 分化制御; 異種生物間遺伝子相補クローニング; iF2α; 細胞周期; Res2; S期開始; 蛋白質リン酸化酵素

During this research term, we have focused on the identification of new elements regulating onset of differentiation of fission yeast and isolated 4 such elements playing key roles regulating the onset of differentiation and switching growth and differentiation. One is phhl^+ encoding a stress MAP kinase highly homologous with mammalian p38. Analysis shows that phhl is required for nutrient starvation-invokes induction of Ste11, a key transcriptional factor essential for the onset of differentiation, providing a molecular basis for the promotion of differentiation by stress. The second is rcdl^+ whose structural homologues are present at least in budding yeast, plant, nematodes and humans. rcdl^+ is essential for nitrogen starvation-invoked differentiation and Ste11 induction. The human homologue of rcdl^+ is expressed abundantly in tests, ovary, spleen and thymus, where cell proliferation and differentiation are actively taking place. The third is nrdl^+ encoding a typical RNA binding … More protein. Analysis shows that the biological role of this gene is to inhibit differentiation by repressing Ste11-regulated genes essential for conjugation and/or meiosis until cells reach a critical point of starvation. We also isolated rat and human homologues of nrdl^+ by expression cloning using fission yeast as host. They are named ROD1. As far as assayd in fission yeast, ROD1 is functionally indistinguishable from nrdl^+. Overexpression of ROD1 in a human hematopietic cell line effectively blocks its differentiation to megakaryocytes.The fourth is srwl^+ encoding a WD repeat protein. Cells lacking this gene are unable to start differentiation, poor to arrestin G1 and defectivein G2 control. The inability to start differentiation is suppressed by deletion of the cig2 cyclin gene, which we previously identified as a negative regulator of differentiation. Analysis shows that srwl^+ is essential for nutrient starvation-induced degradation of the Cdc13 mitotic cyclin. Thus, srwl^+ is a key factor switching between cell proliferation and differentiation. Less

在本研究期间，我们集中于鉴定调控裂殖酵母分化起始的新元件，并分离出4个这样的元件，其在调节分化起始和转换生长和分化中起关键作用。一种是phhl^+，它编码一种与哺乳动物p38高度同源的应激MAP激酶。分析表明，phhl是所需的营养饥饿调用诱导的Ste 11，一个关键的转录因子的分化的开始，提供了一个分子基础，促进分化的压力。第二种是rcdl^+，其结构同源物至少存在于芽殖酵母、植物、线虫和人类中。rcdl^+对于氮饥饿诱导的分化和Ste 11诱导是必不可少的。rcdl^+的人类同源物在睾丸、卵巢、脾脏和胸腺中大量表达，这些部位的细胞增殖和分化活跃。第三个是nrdl^+，它编码一种典型的RNA结合 ...更多信息 蛋白分析表明，该基因的生物学作用是通过抑制Ste 11调节的接合和/或减数分裂所必需的基因来抑制分化，直到细胞达到饥饿的临界点。我们还使用裂殖酵母作为宿主，通过表达克隆分离了nrdl^+的大鼠和人类同源物。它们被命名为ROD 1。就裂变酵母中的检测而言，ROD 1在功能上与nrdl^+没有区别。ROD 1在人造血细胞系中的过表达有效地阻断了其向巨核细胞的分化。第四个是编码WD重复蛋白的srwl^+。缺乏该基因的细胞不能开始分化，对G1抑制差，对G2控制缺陷。不能开始分化是由cig 2细胞周期蛋白基因，我们以前确定为分化的负调节基因的删除抑制。分析表明，srwl^+对于营养饥饿诱导的Cdc 13有丝分裂细胞周期蛋白降解至关重要。因此，srwl^+是在细胞增殖和分化之间转换的关键因子。少

项目成果

Kato,T.Jr.et al.: "Stress signal,mediated by a Hog1-like MAP kinase, controls sexual development in fission yeast." FEBS Let.378. 207-212 (1996)

Kato,T.Jr.等人：“压力信号由 Hog1 样 MAP 激酶介导，控制裂殖酵母的性发育。”

Masuda, A.et al.: "Pole of a signal transduction pathway which dontrols disassembly of microfiament bundles and supperessio of high-molecular weight expression in oncogenic transformation of NRK cells." Oncogene. 12. 2081-2088 (1996)

Masuda, A.等人：“信号转导途径的极点，控制微丝束的分解并抑制 NRK 细胞致癌转化中的高分子量表达。”

Yoshida,S.et al.: "Assembly of Hepatitis C Virus Antigens Expressed from a Recombinant Vaccinia Virus." Proc.Japan Acad.72. 73-78 (1996)

Yoshida,S.et al.：“重组痘苗病毒表达的丙型肝炎病毒抗原的组装”。

Kato, Jr., T., Okazaki, K., Murakami, H.Stettler, S., Fantes, A., P.and Okayama, H.: "Stress signal, mediated by a Hogl-like MAP kinase, controls sexual development in fission yeast." FEBS Let.378. 207-212 (1996)

加藤，Jr.，T.，冈崎，K.，村上，H.Stettler，S.，Fantes，A.，P.和冈山，H.：“压力信号，由 Hogl 样 MAP 激酶介导，控制性行为

Kizaka-Kondoh,S.et al.: "Crkll signals from growth factor receptor to ras." Proc.Natl.Acad.Sci.USA. 93. 12177-12182 (1996)

Kizaka-Kondoh,S.et al.：“Crkll 从生长因子受体向 ras 发出信号。”