BIOCHEMICAL CHARACTERIZATION OF FISSION YEAST KINETOCHORE

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。动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，动粒由至少40个蛋白质亚基组成，形成几个不同的亚复合体。尽管动粒的细胞功能是所有真核生物有丝分裂所普遍需要的，但许多动粒组分并不很保守，至少在一级序列水平上是这样。因此，有必要对多种生物体中的动粒进行研究，以深入了解其分子机制。他的实验室致力于鉴定裂殖酵母中的动粒组分，探索裂殖酵母中动粒的结构以及动粒功能的生化机制。我们采用亲和纯化（TAP）纯化动粒亚复合物的方法，使用特异于鉴定的动粒组分的标签，推定的同源性。到目前为止，我们已经成功地纯化了一种蛋白复合物，对应于芽殖酵母中的DASH复合物，以及另一种可能对应于ctf 3复合物的复合物（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的裂变动粒亚复合物，并进一步研究所确定的蛋白质修饰。

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.