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: 8413853
• 负责人: Tomer Avidor-Reiss
• 金额: $ 27.38万
• 依托单位: UNIVERSITY OF TOLEDO
• 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 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8413853
• 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.

描述（由申请人提供）：中心体由一对中心粒组成，周围环绕着中心粒周围物质（PCM）的无定形蛋白质网络，是对许多细胞功能（包括微管组装和纤毛形成）至关重要的基本细胞组分。有缺陷的中心体生物发生导致男性不育、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，中心粒周围物质（PCM）必须围绕中心粒组装，以使所产生的中心体发挥作用。调节PCM最初如何形成的机制，被招募到中心粒，并围绕中心粒聚集，仍然是一个谜;该提案的目标是解决这个谜。实现这一目标将开始集中在最近发现的细胞质PCM复合物。这种复合物包括多种蛋白质，这些蛋白质在患有小头畸形的患者中是有缺陷的，小头畸形是一种大脑尺寸严重缩小的疾病。本文的目的是分析（1）Sas-4和微管蛋白之间潜在的关键相互作用，（2）Sas-4复合物在纤毛形成中的作用，以及（3）Sas-4复合物在星形微管形成中的作用。这些目标将使用一种跨学科的方法来实现，这种方法将遗传学和生物化学结合在模式生物果蝇中。果蝇具有一些有利的特征，这使其成为研究中心体生物发生的理想工具。首先，许多中心体生物发生突变体是目前可用的。虽然这些突变体中有许多是成虫致死的，但它们不是胚胎或蛹致死的;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，果蝇胚胎可以大量收集，这提供了丰富的参与中心体生物发生的蛋白质来源;这些蛋白质然后可以用于生物化学和体外实验。第三，果蝇的其他众所周知的特征，例如，短世代时间和易于遗传使其成为许多生物系统的首选模式生物，这也与该项目有关。果蝇的这些有利特征应该允许发现PCM形成和组装的机制;这一发现应该有助于开发由缺陷的中心体生物发生引起的人类疾病的诊断和治疗方法。