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Isolation and analysis of new genes that regulate the actin cytoskeleton system

调节肌动蛋白细胞骨架系统的新基因的分离和分析

基本信息

批准号：
11480205
负责人：
TANAKA Kazuma
金额：
$ 7.49万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

The actin cytoskeleton system plays an important role in cell motility, cell-cell adhesion, formation of cell polarity, and cell morphological change. Type I myosins are actindependent motor proteins and are involved in the reorganization of the actin cytoskeleton and endocytosis. Myo3p and Myo5p are the type I myosins in the budding yeast Saccharomyces cerevisiae. In this study, we have isolated a new gene, CDC50, as a multicopy suppressor of the myo5 mutant. The cdc50 mutant was first characterized as a mutant which show cell division cycle arrest, but its functions has not been characterized yet. CDC50 encodes a putative membrane protein that possesses a transmembrane domain. Biochemical experiments indicate that Cdc50p is an integral membrane protein. Microscopic observation of cells expressing CDC50-GFP indicates that CDC50 is localized to endosomes, suggesting that CDC50 is involved in the protein or lipid traffic through endosomes. The cdc50 mutant shows a coldsensitive growth phenotype and cells are arrested at small-budded stage, suggesting that Cdc50p regulates cell polarization. Visualization of the actin cytoskeleton in the cdc50 mutant revealed that actin patches are delocalized in the cdc50 mutant. Our results indicate that CDC50 regulates a traffic to the plasma membrane of a protein that is involved in the polarization of the actin cytoskeleton.

肌动蛋白细胞骨架系统在细胞运动、细胞间粘附、细胞极性形成和细胞形态改变中起着重要作用。I型肌球蛋白是一种不依赖于肌动蛋白的运动蛋白，参与肌动蛋白细胞骨架的重组和内吞作用。Myo 3 p和Myo 5 p是芽殖酵母酿酒酵母中的I型肌球蛋白。在这项研究中，我们已经分离出一个新的基因，CDC 50，作为myo 5突变体的多拷贝抑制。cdc 50突变体首先被鉴定为细胞分裂周期停滞的突变体，但其功能尚未被鉴定。CDC 50编码一种具有跨膜结构域的膜蛋白。生化实验表明Cdc 50 p是一种完整的膜蛋白。对表达CDC 50-GFP的细胞的显微镜观察表明，CDC 50定位于内体，表明CDC 50参与通过内体的蛋白质或脂质运输。cdc 50突变体表现出冷敏感性生长表型，细胞在小出芽阶段被阻滞，表明Cdc 50 p调节细胞极化。在cdc 50突变体的肌动蛋白细胞骨架的可视化显示，肌动蛋白补丁是离域的cdc 50突变体。我们的研究结果表明，CDC 50调节的交通质膜的蛋白质，参与极化的肌动蛋白细胞骨架。