Regulation of Kinetochore Function in Anaphase

Kinetochores mediate interactions between DNA and microtubules during cell division and are essential for proper chromosome segregation. Over 100 human kinetochore components have been identified, but the mechanisms by which these proteins and the kinetochore as a whole adapt their molecular properties to fulfill the changing requirements for chromosome segregation are poorly understood. By comparing the molecular differences between metaphase and anaphase kinetochores and assessing kinetochore functionality by monitoring chromosome movement, I will identify the molecular factors that determine the kinetochore`s ability to segregate sister chromatids. First, I will deplete selected kinetochore proteins to identify the molecular components required for chromosome segregation. Using specialized fusion proteins, I will also eliminate specific proteins acutely in anaphase bypassing their earlier mitotic function and analyze their isolated anaphase function. Second, I will analyze the changes in kinetochore composition and phosphorylation status at anaphase onset and monitor the consequences of disrupting these changes using: (1) specific chemical kinase inhibitors; (2) altered kinase localization and activity; and (3) phosphatase inhibition. My analysis will provide a better understanding of how kinetochores are modified between metaphase and anaphase to segregate the genome accurately.

No additional funding sources recorded.