In vitro phosphorylation of Dam1 complex and Bim1 by Mps1

The Dam1 complex (Dam1c) of Saccharomyces cerevisiae is a major microtubule binding component of the yeast kinetochore. Phosphorylation of various kinetochore proteins by mitotic kinases such as Mps1 are known to modulate the function of these proteins and regulate protein-protein interaction. The Dam1 subunit of Dam1c was previously demonstrated to be phosphorylate by Mps1 (residues S13, S49, S217, S218, S221, S232. In this project we mapped additional phosphor sites on components of the Dam1 complex by mass spectrometry after in vitro phosphorylation as a first step to understand the physiological function of these phosphorylation events.