ROLE OF YEAST MICROTUBULE PROTEIN IN CELL DIVISION

酵母微管蛋白在细胞分裂中的作用

• 批准号: 3081528
• 负责人: DAVID S PELLMAN
• 金额: $ 7.72万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-07-01 至 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3081528
• 关键词: Saccharomyces cerevisiae; binding proteins; cell cycle; cytogenetics; fluorescence microscopy; fungal genetics; gene complementation; gene expression; gene mutation; human tissue; immunofluorescence technique; immunoprecipitation; microtubule associated protein; molecular cloning; nucleic acid sequence; polymerase chain reaction; posttranslational modifications; protein structure function

The regulated assembly and disassembly of microtubule structures is a major aspect of the eucaryotic cell cycle. This dynamic behavior is thought to be regulated in vivo by the action of microtubule-associated proteins (MAPs). Although MAPs have been studied extensively in vitro little is known about their in vivo functions. To be able to understand the role of MAPs in vivo I am studying BIK1, the only characterized non-motor MAP from the yeast, S cerevisiae. The BIK1 protein associates with microtubules in vivo, and this association appears to be restricted to a specific interval in the cell cycle (G2/M). bikl null mutants have phenotypes suggesting that BIK1 plays a role in several aspects of mitotic microtubule function. I am taking two approaches to studying BIK1 function in vivo. The first approach is an analysis of the regions of BIK1 required for association with microtubules and the regions required for the cell cycle dependent change in localization. This will be done by examining the localization of in vitro constructed BIK1 mutants. I will also look for cell cycle dependent modifications of BIK1. The second approach is the analysis of mutants that I have isolated (slb mutants) that require the expression of BIKI for viability, and therefore encode overlapping functions with BIK1. Two of these genes will be cloned, SLB1 and SLB2. I describe experiments to understand the function of BIK1 through an analysis of the physiologic defect of the bik1 slb double mutants. The final goal of this project is to identify BIK1-like genes from human cells and from pathogenic fungi. This will be done by hybridization, or by functional complementation of bik1 slb double mutants. The isolation of such genes would have implications for the development of antineoplastic and antifungal drugs.

微管结构的调节组装和拆卸是一种 真核细胞周期的主要方面。 这种动态行为是 被认为是在体内通过微管相关蛋白的作用进行调节。 蛋白质（MAPs）。 尽管MAPs已经在体外进行了广泛的研究 关于它们在体内的功能知之甚少。 能够理解 MAPs在体内的作用我正在研究BIK 1，唯一表征的 来自酵母，酿酒酵母的非运动MAP。 BIK 1蛋白关联物 与微管在体内，这种协会似乎是有限的 在细胞周期中的特定间隔（G2/M）。bikl无效突变体具有 表型表明BIK 1在以下几个方面发挥作用： 有丝分裂微管功能 我正在采取两种方法来研究BIK 1在体内的功能。 第一 一种方法是分析BIK 1的区域， 与微管和细胞周期依赖的区域 本地化的变化。 这将通过检查 体外构建的BIK 1突变体。 我也会寻找细胞周期 BIK 1的依赖性修饰。 第二种方法是分析 我分离出的突变体（slb突变体）需要表达 BIKI的生存能力，因此编码重叠的功能， 自行车1. 其中两个基因将被克隆，SLB 1和SLB 2。 我描述 通过分析BIK 1的功能， BIK 1 SLB双突变体的生理缺陷。 本项目的最终目标是从人类中鉴定BIK 1样基因， 细胞和致病真菌。 这将通过杂交来完成，或者 通过BIK 1 SLB双突变体的功能互补。 隔离 这些基因的存在可能会影响 抗生素和抗真菌药物。