The Yeast Centrosome - Structure Assembly & Function

酵母中心体 - 结构组装

• 批准号: 8668219
• 负责人: MARK WINEY
• 金额: $ 146.06万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8668219
• 关键词: Address; Alleles; Architecture; Cell Cycle; Cells; Centrosome; Chromosome Segregation; Chromosome abnormality; Chromosomes; Collaborations; Colorado; Complex; Data; Defect; Developmental Delay Disorders; Disease; Environment; Event; Fluorescence Resonance Energy Transfer; Future; Genomic Instability; Goals; Grant; Homeostasis; Human; In Situ; In Vitro; Individual; Knowledge; Lead; Learning; Malignant Neoplasms; Maps; Measures; Mechanics; Microtubule-Organizing Center; Microtubules; Mitosis; Mitotic Chromosome; Mitotic spindle; Modeling; Molecular; Organelles; Principal Investigator; Program Research Project Grants; Property; Proteins; Recruitment Activity; Regulation; Research; Research Personnel; Resources; Roentgen Rays; Rupture; Saccharomyces cerevisiae; Site; Structural Biologist; Structural Models; Structure; Testing; Tubulin; Vertebrates; Washington; Wisconsin; Work; Yeasts; biophysical techniques; chromosome movement; electron tomography; flexibility; in vivo; interdisciplinary approach; interest; metaplastic cell transformation; mutant; nervous system disorder; public health relevance; spindle pole body

DESCRIPTION (provided by applicant): Grant (PPG) will create a collaborative environment to coordinate research on centrosome structure, mechanics, homeostasis and function. Six investigators will study the Saccharomyces cerevisiae centrosome as a model microtubule-organizing center (MTOC) analogous to the vertebrate centrosome, which shares key homologous components and regulators. A centrosome/SPB is the primary microtubule-organizing center of the cell and is critical for bipolar spindle assembly and accurate mitotic chromosome segregation. Centrosome duplication is an essential cell cycle event being the first step in spindle formation; defects in duplication or function lead to genomic instability and cellular transformation. Accurate chromosome segregation depends on both proper regulation of spindle assembly and precise connections between spindle microtubules and chromosomes. The PPG is focused specifically on 10 core SPB components that form the lattice and microtubule nucleation sites including the y-tubulin complexes. These proteins act similarly to the pericentriolar material of vertebrate centrosomes, which is crucial for microtubule nucleation and organization, but poorly understood. We propose to elucidate the molecular architecture of the yeast centrosome and to probe the mechanisms, by which it is assembled, maintained and functions in nucleating microtubules. A multidisciplinary approach examining different aspects of the problem will be coordinated to include: determining how the structure and mechanics of the y-tubulin complex and associated proteins collaborate to accomplish microtubule nucleation; investigating how core SPB components are assembled and how they recruit y-tubulin complexes; identifying the critical intrinsic and extrinsic factors for maintaining homeostasis of this dynamic organelle; solving the atomic structure of SPB components and complexes, and working toward an integrated structural model of the entire SPB core. This PPG builds on existing collaborations between David Agard (UCSF) and Trisha Davis (U. Washington) on g- tubulin complexes, and Ivan Rayment (U. Wisconsin) and Mark Winey (U. Colorado) on core SPB components. These four groups will work together on the 10 proteins. Their projects will profit from structural modeling of the SPB (Andrej Sali, UCSF) and quantifying the mechanical properties of the SPB using biophysical techniques (Chip Asbury, U. Washington). There is tremendous potential to produce an unprecedented molecular description of a centrosome revealing mechanisms of assembly, stability and function. This work will serve as a model for future analysis of the much more complex human centrosome.

描述（由申请人提供）：格兰特（PPG）将创建一个协作环境来协调中心体结构、力学、稳态和功能的研究。六名研究人员将研究酿酒酵母中心体，将其作为类似于脊椎动物中心体的模型微管组织中心（MTOC），其具有关键的同源成分和调节因子。中心体/SPB 是细胞的主要微管组织中心，对于双极纺锤体组装和准确的有丝分裂染色体分离至关重要。中心体复制是一个重要的细胞周期事件，是纺锤体形成的第一步；复制或功能缺陷导致基因组不稳定和细胞转化。准确的染色体分离取决于纺锤体组装的正确调节以及纺锤体微管和染色体之间的精确连接。 PPG 特别关注形成晶格和微管成核位点（包括 y-微管蛋白复合物）的 10 个核心 SPB 组件。这些蛋白质的作用类似于脊椎动物中心体的中心粒周围材料，这对于微管成核和组织至关重要，但人们对此知之甚少。我们建议阐明酵母中心体的分子结构，并探讨其在成核微管中组装、维持和发挥作用的机制。将协调检查问题不同方面的多学科方法，包括：确定 y-微管蛋白复合物和相关蛋白质的结构和力学如何协作以完成微管成核；研究 SPB 核心组件的组装方式以及它们如何招募 y-微管蛋白复合物；确定维持这种动态细胞器稳态的关键内在和外在因素；解决SPB组分和配合物的原子结构，并致力于建立整个SPB核心的集成结构模型。该 PPG 建立在 David Agard（加州大学旧金山分校）和 Trisha Davis（华盛顿大学）在 g-微管蛋白复合物方面的现有合作以及 Ivan Rayment（威斯康星州大学）和 Mark Winey（科罗拉多州大学）在核心 SPB 组件方面的现有合作基础上。这四个小组将共同研究 10 种蛋白质。他们的项目将受益于 SPB 的结构建模（Andrej Sali，加州大学旧金山分校）和使用生物物理技术量化 SPB 的机械性能（Chip Asbury，美国华盛顿州）。对中心体进行前所未有的分子描述具有巨大的潜力，可以揭示组装、稳定性和功能的机制。这项工作将作为未来分析更复杂的人类中心体的模型。