Supplemental Material

This article contains the following supporting material:

• Supplemental Figure - A). pGAL-UBR1-myc cCEN cells were grown in media containing galactose (GAL, inactive cCEN) or glucose (GLU, active cCEN). Cells were harvested for ChIP analysis with anti-myc antibody. Ubr1-myc does not localize to the cCEN or to PGK1. B-C). pGAL-UBR1-myc cCEN cells expressing Okp1-myc13 or Mif2-myc13 and either wild-type CSE4 (+Cse4) or Degron-CSE4 (-Cse4) were grown in media containing galactose and were arrested with nocodazole. Dox was added to repress transcription of Degron-CSE4 and cells were transferred into media containing galactose (GAL, inactive cCEN) or glucose (GLU, active cCEN) and analyzed by ChIP. Vistra-Green quantification is shown. Relative units indicate the cCEN: PGK1 ratio and the amount of epitope-tagged kinetochore protein in GLU +Cse4 is normalized to a value of 1.0. Cse4 is required for full occupancy of Okp1-myc13 and Mif2-myc13. D. Cse4 was depleted from the cCENin cells expressing Stu2-myc13 by incubation with dox for 4 hours. Cells were transferred to media containing galactose (GAL, inactive cCEN) or glucose (GLU, active cCEN) in the presence of dox and then harvested for ChIP. Cse4 is required for the kinetochore localization of Stu2.