Centrosome Structure, Function and Duplication

描述（由申请人提供）：在拟议的研究中解决的核心问题是中心体是如何组织的，它如何在每个细胞周期复制一次，以及我们如何从细胞器的“部件列表”中更全面地了解它是如何工作的。中心体形成微管，并帮助组织这些微管形成有用的阵列，包括有丝分裂纺锤体和纤毛。在过去的20年里，我的实验室和其他人在斯坦福大学的工作已经确定了参与微管成核的分子，中心体复制的中央调节器，参与复制的重要结构蛋白，以及控制中心体数量和连接中心体和纤毛的控制机制。由于中心体异常与癌症发展之间的相关性，以及中心体在纤毛病（与初级纤毛功能相关的疾病）和小头畸形（神经元发育缺陷）中的作用，人们对中心体的兴趣很强。提出的实验利用了我们在不同实验系统中研究中心体结构、功能和重复的试剂和分析方法方面的优势。我们选择关注动物细胞、组织和提取物的过程，因为它们是与我们了解人类正常细胞分裂和疾病中的中心体最相关的系统。本提案涉及三个特定目标：特定目标1 -确定并表征定义中心粒“复制起源”的相互作用；特定目标2 -表征中心粒复制的独特负调节因子Mdm1的作用模式。特异性目标3 -创造无中心粒的哺乳动物细胞，以解决中心体在细胞周期，细胞信号传导和细胞核功能中的功能。

DESCRIPTION (provided by applicant): The central problems addressed in the proposed research are how the centrosome is organized, how it duplicates once per cell cycle, and how we go from the "parts list" for the organelle to a more complete understanding of how it works. The centrosome nucleates microtubules and helps to organize those microtubules to create useful arrays, including the mitotic spindle and the cilium. Work from my lab and others over the last 20 years at Stanford has identified molecules involved in microtubule nucleation, the central regulators of centrosome duplication, important structural proteins involved in duplication, and control mechanisms that govern centrosome number and link the centrosome and the cilium. Interest in the centrosome is strong because of the correlation between centrosome abnormalities and the development of cancer, and because of the role of the centrosome in ciliopathies, diseases related to primary cilium function, and microcephaly, a defect in neuronal development. The proposed experiments make use of the strengths that we have developed in reagents and assays for studying centrosome structure, function and duplication in different experimental systems. We have chosen to focus on the process in animal cells, tissues and extracts as they are the systems most relevant to our desire to understand the human centrosome in normal cell division, and in disease. Three specific aims are addressed in this proposal: Specific Aim 1 - Identify and characterize the interactions that define the centriole "origin of duplication" Specific Aim 2 - Characterize the mode of action of Mdm1, a unique negative regulator of centriole duplication. Specific Aim 3 - Create centriole-less mammalian cells to address the function of the centrosome in the cell cycle, cell signaling, and nuclear functions.