This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Mitotic chromosomal scaffolds contain two multi-protein complexes that are essential for chromosome segregation in mitosis. These are the chromosomal passenger complex formed by Aurora B kinase, INCENP, Survivin and Borealin, and the condensin complex, which contains SMC2, SMC4 and three non-SMC subunits. We intend to further explore the function of these complexes by exploiting DT40 cell lines in which key essential proteins have been conditionally knocked out, and in which the missing component has been complemented by supplying the relevant protein tagged with a tandem affinity tag. This collaboration will involve the use of mass spectrometry methods to identify proteins associated with these complexes in mitotic cells and in mitotic chromosomes. Components thus identified will be confirmed by tagging, production of antibodies, and further subjected to functional analysis. This analysis will identify new cofactors of the chromosomal passenger and condensin complexes, and thereby extend our knowledge of the role(s) of non-histone proteins in regulating chromosomal events of mitosis.
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