STRUCTURE/FUNCTION OF A BASAL BODY-ASSOCIATED CA2+ -BP

基底体相关 CA2 -BP 的结构/功能

• 批准号: 3294175
• 负责人: BESSIE P HUANG
• 金额: $ 23.43万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1993-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3294175
• 关键词: Chlamydomonas; autoradiography; calcium binding protein; cell motility; complementary DNA; computer assisted sequence analysis; cytoskeleton; enzyme linked immunosorbent assay; eukaryote; gel electrophoresis; gene complementation; gene expression; genetic library; genetic mapping; genetic markers; genetic regulation; immunoprecipitation; kinetosome; microtubules; molecular genetics; nucleic acid hybridization; nucleic acid sequence; plant genetics; protein biosynthesis; protein structure function; protein transport; restriction mapping; ultraviolet spectrometry

The long term goals of this research are to understand the molecular mechanisms underlying the biogenesis, segregation, and cellular localization of centers which regulate the assembly and organization of microtubules in eukaryotic cells. The basal body complex in the unicellular green alga Chlamydomonas is used as a model. The specific aims of this proposal are to further characterize the structure/function relationships of a novel basal body-associated 220 kD Ca2+- binding protein (CaBP). During the previous grant period the protein was identified and purified from basal body complexes isolated from Chlamydomonas reihardtii. In studies on the biochemical properties of the purified protein and the characterization of a cDNA containing the entire coding region for the protein, the 20 kD CaBP was found to be clearly related to the calmodulin family of calcium-binding proteins but distinct from well characterized members of this family in its protein sequence, its cellular localization, and potential functions. There are two major features of the 20 kD CaBP which suggest that elucidation of its structure and function may be of fundamental importance in studies on the molecular mechanisms of cell motility and the function and/or disposition of microtubule organizing centers in eukaryotic cells. The 220 kD CaBP has been found to be a component of a calcium-sensitive contractile fiber system which physically links the basal body complex to the nucleus in Chlamydomonas and related algae. In addition to its cytoskeletal localization in algae, evidence has been obtained that sequence-related and antigenically-related proteins to the 20 kD CaBP are associated with the spindle pole body in yeast cells and the centrosome of insight into the structure/function relationship of the 20 kD CaBP as a component of a contractile fiber system which appears to be distinct from actomyosin and microtubles based systems of motility and as a potentially conserved component of microtubule organizing centers of divergent structure in eukaryotic cells.

这项研究的长期目标是了解分子 生物发生、分离和细胞的潜在机制 规范集会和组织的中心的本地化 真核细胞中的微管。基底体复合体位于 使用单细胞绿藻衣藻作为模型。具体的 该提案的目的是进一步描述结构/功能 新型基底体相关 220 kD Ca2+- 结合的关系 蛋白质（CaBP）。在上一次资助期间，该蛋白质是 从分离的基础体复合物中鉴定和纯化 莱氏衣藻。在生化特性的研究中 纯化的蛋白质以及包含整个蛋白质的 cDNA 的表征 蛋白质的编码区，20 kD CaBP 被发现明显是 与钙结合蛋白的钙调蛋白家族相关，但不同 来自该家族的蛋白质序列中已明确表征的成员， 它的细胞定位和潜在功能。主要有两个 20 kD CaBP 的特征表明其 结构和功能在研究中可能具有根本重要性 细胞运动的分子机制以及功能和/或配置 真核细胞中的微管组织中心。 220 kD CaBP 已被发现是钙敏感收缩纤维的组成部分 将基底体复合体与细胞核物理连接的系统 衣藻和相关藻类。除了其细胞骨架外 在藻类中定位，已经获得证据表明序列相关 和 20 kD CaBP 的抗原相关蛋白与 酵母细胞中的纺锤体极体和中心体 20 kD CaBP 作为 a 组件的结构/功能关系 收缩纤维系统似乎与肌动球蛋白不同 基于微管的运动系统并作为潜在的保守系统 不同结构微管组织中心的组成部分 真核细胞。