BIOCHEMICAL CHARACTERIZATION OF FISSION YEAST KINETOCHORE

裂殖酵母动粒的生化特征

• 批准号: 7602235
• 负责人: XIANGWEI HE
• 金额: $ 0.62万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602235
• 关键词: Affinity Chromatography; Biochemical; Cell physiology; Chromosomes; Complex; Computer Retrieval of Information on Scientific Projects Database; Eukaryota; Eukaryotic Cell; Fission Yeast; Funding; Gel; Grant; Institution; Kinetochores; Mediating; Microtubules; Mitosis; Molecular; Organism; Post-Translational Protein Processing; Protein Subunits; Proteins; Research; Research Personnel; Resources; Saccharomycetales; Source; Structure; United States National Institutes of Health; base; interest

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Kinetochore is a highly specialized protein complex assembled on chromosomes that mediates the attachment between chromosome and spindle microtubules. In budding yeast, kinetochores constitute at least 40 protein subunits which form several distinct subcomplexes. Although the cellular functions of the kinetochores are universally required for mitosis in all eukaryotes, many kinetochore components are not well conserved, at least at the primary sequence level. It is therefore necessary to study the kinetochores in multiple organisms for the thorough understanding of their molecular mechanisms. The He lab is interested in identifying the kinetochore components and exploring the kinetochore structure as well as the biochemical mechanism of kinetochore function in fission yeast. We take the approach of purifying kinetochore subcomplexes using affinity purification (TAP), using tags specific to identified kinetochore components, the putative ones based on homology. So far, we have successfully purified a protein complex corresponding to DASH complex in budding yeast and another complex that may correspond to the ctf3 complex (SDS-PAGE gel pictures available upon request). We plan to expand this strategy on more fission kinetochore subcomplexes and also to investigate further the possible protein modifications of the identified ones.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 着丝粒是一种高度特化的蛋白质复合物，组装在染色体上，介导染色体和纺锤体微管之间的附着。在芽殖酵母中，动粒构成至少 40 个蛋白质亚基，这些亚基形成几个不同的亚复合物。尽管着丝粒的细胞功能是所有真核生物有丝分裂所普遍需要的，但许多着丝粒成分并未得到很好的保守，至少在一级序列水平上是如此。因此，有必要研究多种生物体中的着丝粒，以全面了解其分子机制。何实验室的兴趣在于识别裂殖酵母的着丝粒成分并探索着丝粒结构以及着丝粒功能的生化机制。我们采用亲和纯化 (TAP) 来纯化着丝粒子复合物的方法，使用特定于已识别的着丝粒成分的标签，即基于同源性的假定标签。到目前为止，我们已经成功纯化了与芽殖酵母中的 DASH 复合物相对应的蛋白质复合物以及可能与 ctf3 复合物相对应的另一种复合物（可根据要求提供 SDS-PAGE 凝胶图片）。我们计划将这一策略扩展到更多裂变动粒子复合体，并进一步研究已识别的子复合体可能的蛋白质修饰。