MOTOR PROTEINS IN YEAST

酵母中的运动蛋白

• 批准号: 2184230
• 负责人: SUSAN S. BROWN
• 金额: $ 18.04万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2184230
• 关键词: Saccharomyces cerevisiae; calmodulin; cell cycle proteins; chimeric proteins; gene deletion mutation; immunofluorescence technique; immunoprecipitation; interference microscopy; intracellular transport; kinesin; laboratory rabbit; lethal genes; microfilaments; microtubules; myosins; phenotype; phosphorylation; protein structure function; site directed mutagenesis; sporogenesis; suppressor mutations; temperature sensitive mutant; western blottings

An increasing number of both myosins and kinesin-like proteins are being discovered in a given organism, including yeast. The proteins in each of these superfamilies have very similar motor domains, attached to unique domains that may link the motors to different cargos. Yeast is especially well suited for sorting out why there are so many of these motor proteins, and what their separate functions and inter-relationships are. This proposal will study a subset of yeast motor proteins, including two myosins (MYO2p and MYO4p) that show some similarity even outside the motor domain, and a kinesin-like protein (SMY1p) that surprisingly corrects a defect in MYO2p. Several approaches will be used to elucidate their functions: Their motor functions will be tested using in vitro motility assays, and other predicted biochemical properties (calmodulin binding, phosphorylation, and dimerization in the case of the myosins) will be tested for. Antibodies will be made in order to immunolocalize these proteins, and a variety of molecular/genetic techniques will be used (e.g., deletion or overexpression of the genes; screens for suppressors, second-site non-complementors, synthetic lethality; making hybrid proteins or altering them by site-directed mutagenesis).

肌球蛋白和驱动蛋白样蛋白的数量越来越多 在包括酵母在内的给定生物体中发现。 每一个中的蛋白质 这些超家族具有非常相似的电动机域，附在独特 可能将电动机与不同的货物联系起来的域。 尤其是酵母 非常适合整理为什么有这么多的运动蛋白， 以及他们的单独的功能和相互关系。 这 提案将研究酵母运动蛋白的一部分，包括两个肌球蛋白 （myo2p和myo4p）即使在电机域之外也显示出一些相似之处， 和驱动蛋白样蛋白（smy1p），它令人惊讶地纠正了缺陷 myo2p。 将使用几种方法来阐明其功能：他们的功能： 运动功能将使用体外运动测定法和其他 预测的生化特性（钙调蛋白结合，磷酸化和 在肌动物的情况下，将测试二聚化。 抗体 为了使这些蛋白质免疫定位，并将 将使用分子/遗传技术（例如缺失或过表达 基因抑制器，第二站点非汇总者的屏幕， 综合杀伤力；制作混合蛋白或通过 定向诱变）。