Project C

项目C

• 批准号: 7925364
• 负责人: Eva Nogales
• 金额: $ 33.73万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7925364
• 关键词: Anaphase; Aneuploidy; Architecture; BCAS2 gene; Binding; Bioinformatics; Biological Assay; Chromatin; Chromosome Segregation; Chromosomes; Complex; Diffuse; Diffusion; Electron Microscopy; Electrostatics; Elements; Generations; Genetic; Goals; Image Analysis; Kinetochores; Label; Location; Methods; Microtubules; Mitosis; Mitotic; Molecular Probes; Motion; Polymers; Positioning Attribute; Process; Property; Research; Resolution; Role; Site; Structural Models; Structure; Surface; System; Testing; Tubulin; Yeasts; chromosome movement; daughter cell; depolymerization; image reconstruction; protein complex; reconstitution; tumorigenesis

The kinetochore is a network of protein complexes that assembles on centromeric chromatin to act as the connection point between chromosomes and the microtubules that segregate them into daughter cells. During anaphase kinetochores allow chromosomes to track the depolymerizing ends of microtubules, which are the primary site of force generation. Thus, kinetochores not only do not let go of microtubule ends as they breakdown, but are able to harness the energy stored in the microtubule lattice and released during depolymerization to produce processive movement of chromosomes to the spindle poles. In this proposal we aim to probe the molecular mechanisms of directed chromosome motion by examining a set of kinetochore protein complexes that are the site of microtubule attachment. Our approach is to combine activity assays, electron microscopy and image analysis, and bioinformatic methods, in the characterization of fully functional complexes. We are studying the yeast-specific DAM1 complex, which assembles in a microtubule-dependent manner into a ring structure that is able to diffuse on the microtubule surface and to track depolymerizing ends. We are also characterizing the conserved NDC80 complex, and the complete KMN network in yeast, which includes also the MTW1 complex and Spc105. Our ultimate objective is to gain a mechanistic understanding of the molecular interactions governing the dynamic attachment of kinetochores to microtubules. Our structural models would be further tested with our collaborators using the powerful yeast system.

着丝点是一个蛋白质复合物网络，它聚集在着丝染色质上，作为染色体和微管之间的连接点，微管将它们分离成子细胞。在后期，着丝点允许染色体追踪微管的解聚合端，这是产生力的主要部位。因此，着丝点不仅不会在微管末端分解，而且能够利用储存在微管晶格中的能量，并在解聚过程中释放能量，从而使染色体向纺锤极的过程中运动。在这个提议中，我们