The Mechanism of Pericentriolar Material Assembly During Centrosome Biogenesis

中心体生物发生过程中中心粒周围物质组装的机制

• 批准号: 8245269
• 负责人: Tomer Avidor-Reiss
• 金额: $ 20.67万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2012-08-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8245269
• 关键词: Adult; Affect; Animal Model; Animals; Attention; Bardet-Biedl Syndrome; Binding; Binding Sites; Biochemical; Biochemistry; Biogenesis; Biological Assay; Cell division; Cell physiology; Cells; Centrioles; Centrosome; Characteristics; Cilia; Complex; Cystic Kidney Diseases; Cytosol; Data; Defect; Development; Diagnostic; Disease; Drosophila genus; Drosophila melanogaster; Embryo; Eukaryota; Generations; Genetic; Goals; Guanosine Triphosphate; Health; Human; In Vitro; Individual; Left; Male Infertility; Malignant Neoplasms; Mediating; Methods; Microcephaly; Microtubules; Mutation; Neurons; Organelles; Organism; Patients; Positioning Attribute; Process; Proteins; Recruitment Activity; Regulation; Role; Shapes; Source; Structure; Testing; Time; Tubulin; Tubulin Interaction; Work; base; biological systems; brain size; cancer type; cilium biogenesis; developmental disease; experience; fly; in vivo; interdisciplinary approach; mutant; protein complex; protein function; research study; tool

DESCRIPTION (provided by applicant): Centrosomes, which are composed of a pair of centrioles surrounded by an amorphous protein network of pericentriolar material (PCM), are fundamental cellular components that are critical for many cellular functions including microtubule assembly and cilia formation. Defective centrosome biogenesis causes male infertility, numerous types of cancer, and various developmental disorders. Examples of these developmental disorders are cystic kidney disease, Bardet-Biedl syndrome, left-right asymmetry, and Microcephaly. Clearly, proper centrosomes biogenesis is essential to human health. Not surprisingly, centrosome biogenesis is a complex, multi-step process. In particular, pericentriolar material (PCM) must assemble around a centriole for the resulting centrosome to function. The mechanisms regulating how PCM initially forms, is recruited to a centriole, and assembles around the centriole, remain a mystery; the proposal's goal is to solve that mystery. Achieving this goal will begin by focusing on a recently discovered cytoplasmic PCM complex. This complex includes multiple proteins known to be defective in patients suffering from Microcephaly, a disorder where brain size is severely reduced. The aims proposed here are to analyze (1) a potentially key interaction between Sas-4 and tubulin, (2), the role of Sas-4 complexes in cilia formation and (3) the role of Sas-4 complexes in astral microtubule formation. These aims will be accomplished using an interdisciplinary approach that combines genetics and biochemistry in the model organism Drosophila melanogaster. Drosophila possesses several favorable characteristics, which makes it ideal for studying centrosome biogenesis. First, numerous centrosome biogenesis mutants are currently available. Although many of these mutants are adult lethal, they are not embryonic or pupal lethal; thus, Drosophila is one of the only animals in which centrosome biogenesis mutants can be studied in detail. Second, Drosophila embryos can be collected in mass, which provides a plentiful source of the proteins involved in centrosome biogenesis; these proteins can then be used in biochemical and in vitro experiments. Third, other well-known characteristics of Drosophila, e.g., short generation time and ease of genetics, which have made it a preferred model organism for many biological systems, are also relevant to this project. These favorable characteristics of Drosophila should permit discovery of the mechanism of PCM formation and assembly; this discovery should aid in the development of diagnostic and treatment methods for human disorders caused by defective centrosome biogenesis. PUBLIC HEALTH RELEVANCE: Centrosomes, which are composed of a pair of centrioles surrounded by an amorphous protein network of pericentriolar material (PCM), are fundamental cellular components that are critical for many cellular functions. Indeed, defects in centrosomal function are associated with multiple developmental disorders, male infertility, and various forms of cancer. Although much is known about centrosomal structure, the mechanism of PCM formation and assembly around the centrioles remains a mystery.

描述（由申请人提供）：中心体由一对被围叶三环材料（PCM）的无定形蛋白质网络包围的中心元素组成，是对许多细胞功能至关重要的基本细胞成分，包括许多细胞功能，包括微管组装和纤毛形成。中心体生物发生有缺陷会导致男性不育症，多种类型的癌症和各种发育障碍。这些发育障碍的例子是囊性肾脏疾病，Bardet-Biedl综合征，左右不对称和小头畸形。显然，适当的中心体生物发生对人类健康至关重要。毫不奇怪，中心体生物发生是一个复杂的多步骤过程。特别是，周围的三环材料（PCM）必须在中心周围组装，以使其产生的中心体的功能。调节PCM最初形成，招募到中心的机制，并在Centriole周围组装，仍然是一个谜。该提议的目标是解决这个谜。实现这一目标将首先关注最近发现的细胞质PCM复合体。该复合物包括多种蛋白质，已知在患有小头畸形的患者中有缺陷，这种疾病大脑大小严重降低。这里提出的目的是分析（1）SAS-4和微管蛋白之间的潜在关键相互作用，（2），SAS-4复合物在纤毛形成中的作用以及（3）SAS-4复合物在星体微管形成中的作用。这些目标将使用跨学科的方法来实现，该方法结合了模型有机体果蝇中的遗传学和生物化学。果蝇具有多种有利的特征，这是研究中心体生物发生的理想选择。首先，目前有许多中心体生物发生突变体。尽管这些突变体中有许多是成人致死的，但它们不是胚胎或pup的致命。因此，果蝇是唯一可以详细研究中心体生物发生突变体的动物之一。其次，可以在质量中收集果蝇胚胎，从而提供了与中心体生物发生有关的蛋白质的丰富来源。然后，这些蛋白质可用于生化和体外实验。第三，果蝇的其他众所周知的特征，例如短期和遗传学的易感性，这使其成为许多生物系统的首选模型生物，也与该项目有关。果蝇的这些有利特征应允许发现PCM形成和组装的机制；这一发现应有助于开发由中心体生物发生缺陷引起的人类疾病的诊断和治疗方法。 公共卫生相关性：中心体由一对中心元素组成，这些中心元素被围肠里三环材料（PCM）的无定形蛋白质网络（PCM）组成，是对许多细胞功能至关重要的基本细胞成分。实际上，中心体功能的缺陷与多种发育障碍，男性不育症和各种形式的癌症有关。尽管对中心体结构知之甚少，但围绕中心的PCM形成和组装的机理仍然是一个谜。