CELLULAR FUNCTION OF ACTIN HOMOLOGUES

肌动蛋白同系物的细胞功能

• 批准号: 2186912
• 负责人: DAVID I MEYER
• 金额: $ 16.4万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2186912
• 关键词: actin binding protein; cell cycle; centrosome; density gradient ultracentrifugation; gel electrophoresis; gene complementation; gene expression; immunocytochemistry; immunofluorescence technique; immunoprecipitation; laboratory mouse; laboratory rabbit; microinjections; nucleic acid sequence; phenotype; polymerase chain reaction; polymerization; posttranslational modifications; protein isoforms; protein structure function; southern blotting; tissue /cell culture; yeasts

Centractins represent a family of highly-conserved mammalian genes whose sequences indicate a significant degree of similarity (>70%) to actin. Members of this family have been localized to centrosomes and large vesicular structures by morphological analyses. In addition, biochemical studies have shown that centractins are associated with a 20S cytosolic proteinaceous particle known as the p150(Glued), or dynactin complex. This complex has been implicated in the function of cytoplasmic dynein, a molecular motor which is responsible for retrograde vesicular transport along microtubules. As at least two isoforms of centractin have been characterized in our laboratory, as well as a novel yeast actin-related protein (ARP) which may represent a functional homologue of centractin. It is the aim of the research described here to use biochemical, morphological and genetic techniques to answer the most fundamental questions pertinent to the role of centractins in cellular function. These include: As centractin isoforms have individual subcellular localizations, how do they differ functionally? Are centractins capable of polymerizing into multimeric complexes or filaments, in a fashion homologous to actin, or to serve as actin-binding proteins? What is the "centractin phenotype" in mammalian cells whose centractin expression or integrity has been compromised? Is the yeast ARP, ACT3, the functional homologue of centractin, and if so what is its role in this organism?

中心蛋白代表了一个持持牢固的哺乳动物基因的家族 序列表明与肌动蛋白相似程度很高（> 70％）。 这个家庭的成员本地化为中心体和大型 囊泡结构通过形态分析。另外，生化 研究表明，中心蛋白与20s胞质有关 蛋白质颗粒称为p150（胶合）或dynactin复合物。这 复合物与细胞质动力素的功能有关 分子电动机，负责逆行囊泡转运 沿着微管。至少有两个中心蛋白的同工型 在我们的实验室以及新型酵母肌动蛋白相关的特征 蛋白质（ARP）可能代表中心乳蛋白的功能同源物。 这里描述的研究目的是使用生化， 形态学和遗传技术回答最基本的 与中心素在细胞功能中的作用有关的问题。这些 包括：由于中心乳蛋白同工型具有个体的亚细胞局部局部化，因此 它们的功能有何不同？中心素能够聚合 以与肌动蛋白同源的方式进入多聚体络合物或细丝 还是用作肌动蛋白结合蛋白？什么是“中心元表型” 在中心素表达或完整性的哺乳动物细胞中 妥协？是酵母ARP，ACT3，功能同源 中心蛋白，如果是的，它在这种生物体中的作用是什么？