BIOCHEMICAL CHARACTERIZATION OF FISSION YEAST KINETOCHORE

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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.