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: 8643261
• 负责人: Tomer Avidor-Reiss
• 金额: $ 24.84万
• 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/8643261
• 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，中心粒被中心粒周围物质（PCM）的无定形蛋白质网络包围，是细胞的基本成分，对许多细胞功能至关重要，包括微管组装和纤毛形成。中心体生物发生缺陷导致男性不育、多种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，正确的中心体生物形成对人类健康至关重要。不出所料，中心体的生物发生是一个复杂的、多步骤的过程。特别是，中心粒周围物质（PCM）必须聚集在中心粒周围，从而使中心体发挥作用。调控PCM最初形成、被招募到中心粒并在中心粒周围组装的机制仍然是一个谜；该提案的目标就是解开这个谜团。实现这一目标将从关注最近发现的细胞质PCM复合物开始。这种复合体包括多种已知在患有小头症的患者中存在缺陷的蛋白质，小头症是一种大脑大小严重缩小的疾病。本文的目的是分析(1)Sas-4与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；(3)Sas-4复合物在星状微管形成中的作用。这些目标将完成使用跨学科的方法，结合遗传学和生物化学的模式生物黑腹果蝇。果蝇具有几个有利的特征，使其成为研究中心体生物发生的理想动物。首先，目前有许多中心体生物发生突变体。虽然这些突变体在成年时是致命的，但它们在胚胎或蛹时不是致命的；因此，果蝇是唯一一种可以详细研究中心体生物发生突变的动物。其次，可以大量收集果蝇胚胎，这为中心体生物发生提供了丰富的蛋白质来源；这些蛋白质可以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，短的繁殖时间和易于遗传，使其成为许多生物系统的首选模式生物，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；这一发现将有助于开发诊断和治疗由中心体生物发生缺陷引起的人类疾病的方法。