REPLICATION AND SEGREGATION OF BASAL BODIES

基础体的复制和分离

• 批准号: 2190828
• 负责人: CAROLYN D SILFLOW
• 金额: $ 19.94万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-02-01 至 1999-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2190828
• 关键词: Chlamydomonas; cell cycle; cilium /flagellum motility; flagellum; gene expression; gene mutation; immunocytochemistry; kinetosome; molecular cloning; nucleic acid sequence; transposon /insertion element

Cilia and flagella, and the basal bodies from which they grow, are highly conserved throughout the evolution of eukaryotic cells. from unicellular protists to various types of differentiated cells in mammalian systems. Basal bodies, which are analogous to centrioles in structure, serve as the organizing site for a complex set of cytoskeletal structures including microtubule rootlets and various striated fibers. These cytoskeletal elements appear to be involved in segregating basal bodies properly during the cell cycle, in determining or maintaining cellular polarity, in positioning of other cellular organelles, and in anchoring cilia and flagella in the cell. The unicellular biflagellate green alga Chlamydomonas has long been used as a model genetic system to study the genes involved in flagellar function. Previous genetic studies of mutants with abnormal flagellar number have shown that this phenotype results from defects in the replication, maturation, and segregation of basal bodies during the cell cycle. The recent development of methods for insertional mutagenesis in Chlamydomonas has greatly facilitated the cloning of genes identified by mutation. These new methods will be used to clone genes involved in basal body function. The specific aims are: 1) to use insertional mutagenesis to identify genes involved in basal body replication, segregation, and localization by isolating mutants with abnormal flagellar number; 2) to clone the genes identified by mutation and to characterize them by DNA sequencing; 3) to examine the function of each gene through detailed cytological analysis of the mutant phenotype; and 4) to examine the spatial and temporal localization of the gene products using immunocytochemistry. It is likely that many of the proteins involved in basal body function identified in this project will have homologs in mammalian cells such as ciliated epithelial cells, sperm cells, and sensory cells. Results from this project will provide a better understanding of the cytoskeleton in these cells.

纤毛和鞭毛，以及它们生长的基体， 在真核细胞的进化过程中是保守的。从单细胞 原生生物到哺乳动物系统中各种类型的分化细胞。 在结构上类似于中心粒的基体， 一组复杂的细胞骨架结构的组织部位，包括 微管根和各种条纹纤维。 这些细胞骨架 元素似乎参与分离基体适当期间 细胞周期，决定或维持细胞极性， 定位其他细胞器，并在锚定纤毛和 细胞中的鞭毛 单细胞双鞭毛绿色藻 衣原体长期以来一直被用作模型遗传系统来研究 参与鞭毛功能的基因 以前的突变体遗传研究 异常鞭毛数量的研究表明，这种表型是由 基体复制、成熟和分离方面的缺陷 在细胞周期中。 本文综述了近年来插入法的研究进展 衣原体的突变极大地促进了基因的克隆 通过突变识别。 这些新方法将用于克隆基因 参与身体的基本功能。 具体目标是：1）使用 插入突变以鉴定涉及基体的基因 复制，分离和定位，通过分离突变体， 2）克隆突变鉴定的基因 并通过DNA测序对其进行表征; 3）检测 通过详细的细胞学分析每个基因的突变表型; 以及4）检查基因的时空定位 产品使用免疫细胞化学。 很有可能， 本项目中确定的参与基础身体功能的蛋白质将 在哺乳动物细胞中有同源物，如纤毛上皮细胞、精子 细胞和感觉细胞。 该项目的结果将提供更好的 了解这些细胞中的细胞骨架。