Mechanisms of centriole assembly and stability

描述（由申请人提供）：中心粒是成核和组织中心体和纤毛的细胞纳米机器。中心粒缺陷会导致癌症和纤毛病变。纤毛病是一种常见的人类疾病，包括儿童出生缺陷、智力迟钝、多指、失明、肥胖和多囊性肾。然而，中心粒形成的机制，当有缺陷时，有助于上述病理还没有很好地理解。我们的长期目标是了解这些缺陷的病因。为了实现这一目标，我们将研究中心粒形成和功能中的两个重要事件：组装和稳定。这两个事件对中心体和纤毛都至关重要。这两个事件都需要一个基本的中心粒结构，那就是侧手翻。为了阐明侧手翻的分子组装，将采用生物化学、分子遗传学和定量成像策略。首先，我们将确定一个新的复杂的侧手翻组件稳定中心粒。其次，我们将定义蛋白质组分如何以及何时相互作用以构建侧手翻和中心粒结构。第三，稳定复合物的一个成员是Bld 10/Cep 135，除了其稳定作用外，它对中心粒组装至关重要。Bld 10在中心粒功能中的作用将通过分离其在中心粒组装和维护中的功能并鉴定与其相互作用以执行这些功能的蛋白质来确定。我们将建立车轮和中心粒组装成一个稳定结构的关键蛋白质和机制。在这个稳定结构的基础上，我们可以定义这个平台如何使纤毛和中心体成核。这项建议将发展我们的长期目标，以重建侧手翻和中心粒组装，并了解这些过程中的缺陷如何导致人类疾病。 我们的研究将由一个研究人员合作小组进行。这些研究将为本科生和研究生以及博士后研究人员和专业研究助理提供出色的培训经验。与包括计算建模，定量图像分析和蛋白质组学在内的其他实验室的合作将扩大我们研究的影响。

DESCRIPTION (provided by applicant): Centrioles are cellular nanomachines that nucleate and organize centrosomes and cilia. Defects in centrioles contribute to both cancer and ciliopathies. Ciliopathies are a general class of human maladies that include child birth defects, mental retardation, polydactyl, blindness, obesity, and polycystic kidneys. However, the mechanics of centriole formation that, when defective, contribute to the above pathologies are not well understood. Our long term goals are to understand the etiology of such defects. Toward this goal, we will study two important events in centriole formation and function; assembly and stabilization. Both of these events are vital for centrosomes and cilia. An essential centriole structure that is required for both of these events is the cartwheel. To elucidate the molecular assembly of a cartwheel, biochemical, molecular-genetic, and quantitative imaging tactics will be employed. First, we will determine how a novel complex of cartwheel components stabilizes centrioles. Second, we will define how and when protein components interact to build the cartwheel and centriole architecture. Third, one member of the stability complex is Bld10/Cep135 that, in addition to its stability role, is essential for centriole assembly. Bld10's roles in centriole function will be determined by separating its functions in centriole assembly and maintenance and identifying the proteins it interacts with to perform these functions. We will establish the key proteins and the mechanisms for cartwheel and centriole assembly into a stable structure. Building on this stable structure, we can then define how this platform nucleates cilia and centrosomes. This proposal will develop our long term goals to reconstitute cartwheel and centriole assembly and to understand how defects in these processes cause human disease. Our studies will be carried out by a collaborative group of researchers. These studies will provide excellent training experiences for undergraduate and graduate students and postdoctoral researchers and professional research assistants. Collaborations with other labs that include computational modeling, quantitative image analysis, and proteomics will expand the impact of our studies.