The Role of CP110 in Ciliogenesis

CP110 在纤毛发生中的作用

• 批准号: 8663546
• 负责人: Brian D Dynlacht
• 金额: $ 16.78万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8663546
• 关键词: Ablation; Affinity Chromatography; Animal Model; Biochemical; Biochemistry; Carrier Proteins; Cell Cycle Progression; Cell membrane; Cell surface; Cellular biology; Centrioles; Centrosome; Cilia; Coatomer Protein; Complex; Congenital Abnormality; Defect; Development; Disease; Docking; Exhibits; Genes; Golgi Apparatus; Human; Laboratories; Lead; Malignant Neoplasms; Mammalian Cell; Mammals; Maps; Mass Spectrum Analysis; Mitotic spindle; Molecular; Monitor; Mutation; Organelles; Orthologous Gene; Phenotype; Play; Process; Proteins; Proteomics; RNA Interference; Role; Sensory; Structure of ciliary processes; Testing; Vesicle; Work; base; cell growth; ciliopathy; cilium biogenesis; fly; interest; kinetosome; mutant; novel; protein function; protein transport; receptor; retrograde transport; trafficking

DESCRIPTION (provided by applicant): Our work seeks to understand the controls that regulate assembly and organization of the mitotic spindle and cilia assembly. Both of these processes are governed by the centrosome, an organelle that remains largely enigmatic. We are particularly interested in elucidating the molecular mechanisms that govern the switch between centriolar and basal body function, as the assembly of cilia from basal bodies is vital for a large number of developmental decisions. Many human birth defects, developmental abnormalities, and ciliopathies result from defective ciliogenesis or mutations in ciliary proteins. Recently, we have identified and characterized a centrosomal protein, CP110, and a cadre of associated proteins that play important roles in centrosome duplication, assembly of the mitotic spindle, and ciliogenesis. Ultimately, we hope to understand how this network of proteins regulates primary cilia assembly and function and how this process is disturbed in human developmental diseases. Recently, we identified several novel proteins that interact with CP110 and Cep97, including Talpid3 and WDR35, and we have begun to characterize the role of each protein in the context of cilia assembly. Mutations in Talpid3 and WDR35 give rise to developmental defects in diverse animal models, and mutations in WDR35 give rise to birth defects in humans. However, the mechanisms underlying the function of these proteins are not known. Our studies indicate that both proteins are involved in cilia assembly and function. Our proposal is based on three aims that use a combination of biochemistry, proteomics, cell biology, and RNAi. First, we will investigate the function of Talpid3 and WDR35 and ask how these proteins interact with the CP110 complex and how they direct cilia assembly. Next, we will ask how defects in these proteins lead to ciliary defects. We will investigate whether Talpid3 and WDR35 play roles in protein transport between vesicles and basal bodies/cilia and whether they regulate cell growth and quiescence. Third, we will perform biochemical screens to identify proteins that interact with Talpid3 and WDR35. In addition to explaining basic biochemical details regarding the role of CP110-associated proteins, trafficking of proteins into and out of the cilium, and ciliogenesis-processes that remain poorly understood--we will begin to understand the mechanisms underlying human ciliopathies and birth defects resulting from mutations in ciliary disease genes and the phenotypes associated with these diseases.

描述(由申请人提供)：我们的工作试图了解调控有丝分裂纺锤体和纤毛组装的组装和组织的控制。这两个过程都是由中心体控制的，中心体是一个在很大程度上仍然神秘的细胞器。我们特别感兴趣的是阐明控制中心粒和基本体功能切换的分子机制，因为来自基本体的纤毛组装对于大量的发育决定是至关重要的。许多人类先天缺陷、发育异常和纤毛疾病都是由纤毛发生缺陷或纤毛蛋白突变引起的。最近，我们已经鉴定了中心体蛋白CP110和一系列相关蛋白，它们在中心体复制、有丝分裂纺锤体的组装和纤毛发生中发挥重要作用。最终，我们希望了解这一蛋白质网络如何调节初级纤毛的组装和功能，以及这一过程在人类发育疾病中是如何受到干扰的。最近，我们发现了几种新的与CP110和Cep97相互作用的蛋白质，包括Talpid3和WDR35，我们已经开始研究每种蛋白质在纤毛组装中的作用。Talpid3和WDR35的突变会导致不同动物模型的发育缺陷，而WDR35的突变会导致人类的出生缺陷。然而，这些蛋白质潜在的功能机制尚不清楚。我们的研究表明，这两种蛋白都参与了纤毛的组装和功能。我们的建议基于三个目标，结合使用生物化学、蛋白质组学、细胞生物学和RNAi。首先，我们将研究Talpid3和WDR35的功能，并询问这些蛋白质如何与CP110复合体相互作用，以及它们如何指导纤毛组装。接下来，我们将询问这些蛋白质的缺陷是如何导致纤毛缺陷的。我们将研究Talpid3和WDR35是否在囊泡和基底体/纤毛之间的蛋白质运输中发挥作用，以及它们是否调节细胞的生长和静止。第三，我们将进行生化筛选，以确定与Talpid3和WDR35相互作用的蛋白质。除了解释CP110相关蛋白的作用、蛋白质进出纤毛和纤毛发生的基本生化细节-这些过程仍然知之甚少--我们还将开始了解人类纤毛疾病和出生缺陷的机制，这些机制是由睫状结构疾病基因突变引起的，以及与这些疾病相关的表型。