BASAL BODY DUPLICATION IN CHLAMYDOMONAS

衣藻的基础体复制

• 批准号: 8362539
• 负责人: SUSAN K DUTCHER
• 金额: $ 1.06万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362539
• 关键词: Cell Cycle; Cell Growth Processes; Cell physiology; Cells; Cellular Structures; Centrioles; Chlamydomonas; Complex; Cryoultramicrotomy; Freezing; Funding; Grant; Image; Light; Messenger RNA; Microtubules; Mitotic; National Center for Research Resources; Paramecium; Plastics; Principal Investigator; Proteins; RNA Interference; Research; Research Infrastructure; Resources; Source; Tomogram; United States National Institutes of Health; base; cost; 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. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 关于基础体和中心元素的组装知之甚少。 为了更多地了解重复过程，可以通过光与黑暗的周期同步细胞的生长和分裂周期，以检查基础体积重复的早期步骤。 此外，我们有两个突变体可能特别感兴趣。第一个是一种菌株，该菌株具有存在于野生型基体底部的无定形环，但突变基底体不组装微管。因此，它的表征对于早期步骤很重要。 第二个是RNA干扰菌株，该菌株的mRNA显着降低了富含Luecine的重复蛋白。它产生了太多的基体。 该蛋白质似乎是基底体/中心重复的负调节剂，并且在基础体内隔离中没有缺陷。 同步菌株将使用高压冷冻并冻结塑造并嵌入塑料的材料来表征，以重建整个基底体络合物，以串行断层造影重建。迄今为止，已经成像了4个有丝分裂细胞的串行断层图，以揭示出发展概率的体。这些特征在于单线微管的环，类似于Pragecium中描述的环。此外，还将切割冷冻水合材料的冷冻切片，并将成像冷冻图图，以获取复制中间体的详细信息，以接近天然状态的方式。