Figure 4.

Cse4 is required for the full occupancy of inner and some central kinetochore components. (A–F). pGAL-UBR1-myc cCEN cells expressing the indicated myc-epitope tagged kinetochore protein and either wild-type CSE4 (+Cse4) or Degron-CSE4 (–Cse4) were grown in media containing galactose and arrested with nocodazole. Dox was added to repress transcription of Degron-CSE4, and cells were transferred into media containing GAL (inactive cCEN) or GLU (active cCEN) and analyzed by ChIP with anti-myc antibody. (E and F) Quantification of A and C. Relative units indicate the cCEN:PGK1 ratio. The cCEN:PGK1 value in GLU +Cse4 is normalized to a value of 1.0. Cse4 is required for the complete occupancy of Ndc10-myc13, Mif2-myc13, Mtw1-myc13, and Okp1-myc13 (also see Supplemental Figure 1, B and C).