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。

基本信息

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

项目摘要

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，我们重点鉴定了调节裂殖酵母分化开始的新元件，并分离出 4 个在调节分化开始以及转换生长和分化方面发挥关键作用的元件，其中之一是编码应激 MAP 激酶的 phl^+。分析表明，phl 是营养饥饿所必需的——诱导 Ste11，这是分化开始所必需的关键转录因子，为通过应激促进分化提供了分子基础。 rcdl^+ 的结构同源物至少存在于芽殖酵母、植物、线虫和人类中。rcdl^+ 对于氮饥饿引起的分化和 Ste11 诱导至关重要。 rcdl^+ 的人类同源物在测试、卵巢、脾脏和胸腺，细胞增殖和分化活跃地发生。第三个是 nrdl^+，编码一种典型的 RNA 结合蛋白。分析表明其生物学作用。该基因通过抑制接合和/或减数分裂所必需的 Ste11 调节基因来抑制分化，直到细胞达到饥饿的临界点。我们还通过使用裂殖酵母作为宿主进行表达克隆，分离了 nrdl^+ 的大鼠和人类同源物。就裂殖酵母中的检测而言，ROD1 在功能上与人类造血细胞系中的 nrdl^+ 没有区别。第四个是 srwl^+，编码 WD 重复蛋白。缺乏该基因的细胞无法开始分化，抑制蛋白 G1 和 G2 控制缺陷。通过删除 cig2 细胞周期蛋白基因，可以抑制无法开始分化。，我们之前将其确定为分化的负调节因子。分析表明，srwl^+ 对于营养饥饿诱导的 Cdc13 有丝分裂细胞周期蛋白的降解至关重要。 srwl^+是细胞增殖和分化之间切换的关键因素。