Genetic analysis of basal body function

身体基础功能的遗传分析

基本信息

批准号：
7924929
负责人：
SUSAN K DUTCHER
金额：
$ 19万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7924929
关键词：
Address Algae Animals Antibodies Back Biological Biological Assay Candidate Disease Gene Cell Cycle Cell Nucleus Cell division Cells Centrioles Centrosome Chlamydomonas Chromosome Mapping Chromosome Segregation Cilia Collection Daughter Dynein ATPase Enhancers Event Family member Flagella Genes Genetic Genome Stability Genomics Immigration Immunoelectron Microscopy Immunofluorescence Immunologic Interphase Cell Investigation Knowledge Laboratories Left Maps Mature Centriole Microtubule-Organizing Center Microtubules Mining Mitosis Mitotic Modeling Molecular Motors Mothers Mutation Organelles Organism Pathway interactions Protein phosphatase Proteins RNA Interference Reporter Reporter Genes Role Staining method Stains Structure Testing Time Tubulin aurora-A kinase cell type comparative gamma Tubulin genetic analysis kinetosome migration mutant trait tumor progression

项目摘要

DESCRIPTION (provided by applicant): In animal cells microtubules are organized by the centrosome, an organelle with a pair of centrioles enclosed in a matrix ofpericentriolar material. Once per cell cycle, the centrosome and centrioles within it duplicate. This event is critical to cell division and genome stability. Duplication involves the separation of the existing centriole pair and the synthesis of a new centriole adjacent to the old centriole. The new pairs of centrioles migrate apart to the poles of the assembling spindle. Centrioles can mature into basal bodies that template flagella and cilia. The unicellular alga, Chlamydomonas provides an outstanding model for investigations into centrioles/basal bodies because of its straightforward cell biological traits and powerful genetics. In the last few years, studies from my laboratory have helped to demonstrate the complexity of basal bodies and to identify critically important roles for two new tubulin family members in centriole/basal body duplication and function. These studies have left unanswered several important questions and we will address three of these questions: 1). What are the molecules needed for early events in centriole duplication? 2). What are the molecules involved in maturation of centrioles through the cell cycle? 3). How do centrioles separate and migrate at mitosis? We will employ mutations, antibodies, and RNA interference in Chlamydomonas to answer these questions. We will test the roles of epsilon- and eta-tubulin in centriole duplication as well as the roles of several suppressors and enhancers of epsilon-tubulin mutations. We hypothesize that at least four genes, delta- tubulin, DT11, DT12, and UN13, are needed to mature centrioles into basal bodies at the M/G1 transition. We will develop antibody markers for maturation and begin to characterize the events and players in maturation. Basal bodies are converted back into centrioles that separate, migrate, and organize events in preprophase of mitosis. Using RNA interference, we will test the roles of several proteins that are important in other organisms and with this knowledge screen for conditional mutants using a GFP-gamma-tubulin reporter to follow centriole separation and migration. We will mine using "comparative genomics" to identify additional genes that are needed for duplication, maturation, and migration of the centrioles/basal bodies.

描述（由申请人提供）：在动物细胞中，微管由中心体组织而成，中心体是一种细胞器，具有一对中心粒，包围在中心粒周围材料的基质中。每个细胞周期，中心体和其中的中心粒复制一次。该事件对于细胞分裂和基因组稳定性至关重要。复制涉及现有中心粒对的分离以及与旧中心粒相邻的新中心粒的合成。新的中心粒对分开迁移到装配轴的两极。中心粒可以成熟为模板鞭毛和纤毛的基体。单细胞藻类衣藻因其简单的细胞生物学特性和强大的遗传学特性，为研究中心粒/基体提供了一个出色的模型。在过去的几年中，我实验室的研究帮助证明了基底体的复杂性，并确定了两个新的微管蛋白家族成员在中心粒/基底体复制和功能中至关重要的作用。这些研究没有回答几个重要问题，我们将解决其中三个问题：1）。中心粒复制的早期事件需要哪些分子？ 2）。细胞周期中参与中心粒成熟的分子有哪些？ 3）。有丝分裂时中心粒如何分离和迁移？我们将利用衣藻突变、抗体和 RNA 干扰来回答这些问题。我们将测试ε-微管蛋白和eta-微管蛋白在中心粒复制中的作用，以及几种ε-微管蛋白突变抑制子和增强子的作用。我们假设至少需要四个基因：δ-微管蛋白、DT11、DT12 和 UN13，才能在 M/G1 转变时使中心粒成熟为基体。我们将开发成熟的抗体标记，并开始描述成熟过程中的事件和参与者。基体转变回中心粒，在有丝分裂前期分离、迁移和组织事件。使用 RNA 干扰，我们将测试几种在其他生物体中重要的蛋白质的作用，并利用这一知识，使用 GFP-γ-微管蛋白报告基因筛选条件突变体，以跟踪中心粒分离和迁移。我们将使用“比较基因组学”进行挖掘，以确定中心粒/基体复制、成熟和迁移所需的其他基因。