BASAL BODY DUPLICATION IN CHLAMYDOMONAS

衣藻的基础体复制

• 批准号: 8170837
• 负责人: SUSAN K DUTCHER
• 金额: $ 2.49万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170837
• 关键词: Cell Growth Processes; Cells; Centrioles; Chlamydomonas; Complex; Computer Retrieval of Information on Scientific Projects Database; Cryoultramicrotomy; Freezing; Funding; Grant; Image; Institution; Light; Messenger RNA; Microtubules; Mitotic; Paramecium; Plastics; Proteins; RNA Interference; Research; Research Personnel; Resources; Source; Tomogram; United States National Institutes of Health; base; interest; kinetosome; mutant; pressure; reconstruction; segregation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Little is known about the assembly of basal bodies and centrioles. To understand more about the duplication process, the cell growth and division cycle can be synchronized by cycles of light and dark to examine the early steps in basal body duplication. In addition, we have two mutants that may be of particular interest. The first is a strain that has the amorphous ring that is present at the base of wild-type basal bodies, but the mutant basal bodies do not assemble microtubules. Thus, its characterization is important to look at early steps. The second is an RNA interference strain that has significant reduction in the mRNA for a luecine rich repeat protein; it makes too many basal bodies. This protein appears to act as a negative regulator of basal body/centriole duplication and is not defective in basal body segregation. Synchronized strains will be characterized using material that has been high pressure frozen and freeze-substituted and embedded in plastic to reconstruct the entire basal body complex in serial tomographic reconstructions. To date, 4 serial tomograms of mitotic cells have been imaged that reveal developing probasal bodies. These are characterized by rings of singlet microtubules, similar to what has been described in Paramecium. In addition, cryosections of frozen hydrated material will also be cut and cryotomograms will be imaged to get detailed information of duplication intermediates in a manner close to the native state.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 关于基体和中心粒的组装知之甚少。 为了更多地了解复制过程，可以通过光和暗周期使细胞生长和分裂周期同步，以检查基体复制的早期步骤。 此外，我们还有两个可能特别令人感兴趣的突变体。第一种是具有存在于野生型基体基部的无定形环的菌株，但突变型基体不组装微管。因此，其特征对于研究早期步骤非常重要。 第二种是 RNA 干扰菌株，其富含亮氨酸的重复蛋白的 mRNA 显着减少；它产生了太多的基体。 该蛋白质似乎充当基体/中心粒复制的负调节因子，并且在基体分离方面没有缺陷。 将使用高压冷冻和冷冻替代并嵌入塑料中的材料来表征同步菌株，以在连续断层扫描重建中重建整个基体复合体。迄今为止，已对有丝分裂细胞的 4 个系列断层扫描图像进行成像，显示正在发育的基体。它们的特征是单线态微管环，类似于草履虫中所描述的。此外，还将切割冷冻水合材料的冷冻切片并对冷冻断层图进行成像，以接近天然状态的方式获得重复中间体的详细信息。