BIOCHEMICAL CHARACTERIZATION OF FISSION YEAST KINETOCHORE

裂殖酵母动粒的生化特征

• 批准号: 7420715
• 负责人: XIANGWEI HE
• 金额: $ 0.29万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7420715
• 关键词: centromere; proteins; yeasts

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.

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。动粒是一种组装在染色体上的高度专门化的蛋白质复合体，它介导染色体和纺锤体微管之间的附着。在萌芽酵母中，动粒至少由40个蛋白质亚基组成，形成几个不同的亚复合体。虽然动粒的细胞功能在所有真核生物中都是有丝分裂所必需的，但许多动粒成分并不是很保守，至少在初级序列水平上是这样。因此，有必要对多种生物中的动点进行研究，以深入了解它们的分子机制。该实验室感兴趣的是鉴定动粒成分，探索动粒结构以及分裂酵母中动粒功能的生化机制。我们采用亲和纯化(TAP)的方法来纯化动粒亚复合体，使用识别的动粒成分的特异标签，这些成分是基于同源性的假定成分。到目前为止，我们已经成功地纯化了一个与发芽酵母中的DASH复合体相对应的蛋白质复合体，以及另一个可能与ctf3复合体相对应的复合体(可根据需要获得SDS-PAGE凝胶图片)。我们计划将这一策略扩展到更多的分裂动粒亚复合体上，并进一步研究已鉴定的亚复合体的可能的蛋白质修饰。