Abstract
We are interested in understanding how centromeres are specified and how centromeric chromatin forms. To this end, we immunopurified the centromeric histone H3 variant Cse4 from budding yeast and subjected this preparation to MudPIT analysis to identify interacting proteins. Peptides of Scm3 were identified. Scm3 is essential for cell viability and localizes to all centromeres. Construction of a conditional SCM3 allele reveals that depletion results in metaphase arrest, with duplicated spindle poles, short spindles, and unequal DNA distribution. The metaphase arrest is mediated by the mitotic spindle checkpoint, being dependent on Mad1 and the Aurora kinase B homolog Ipl1. Scm3 interacts with both Ndc10 and Cse4 and is essential to establish centromeric chromatin following DNA replication. In addition, Scm3 is required to maintain kinetochore function throughout the cell cycle. This research is supported by the Stowers Institute for Medical Research.
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