Figure 5.

Overexpression of Cse4p does not phenocopy loss of Psh1p in increasing CEN plasmid protein levels. (A) CHX chase for the indicated times assessing turnover of myc-Cse4p in WT and psh1Δ yeast cells. Proteins were detecting by immunoblotting with myc antibody. Both long and short exposures are shown. Anti-PGK serves as a loading control. (B) The steady-state protein levels of CEN plasmid-expressed mia40-4pHA and Fzo1pHA, as well as endogenous genome-expressed Cse4p, were detected by immunoblotting with HA or Cse4p antibodies, in the indicated deletion strains. (C) Immunoblotting for Cse4p using Cse4p antibodies in WT cells, psh1Δ cells, or cells overexpressing Cse4p from the genome (pGAL-FLAG-CSE4) for the indicated time points after continuous galactose induction. (D) Immunoblotting using anti-HA for CEN plasmid-expressed mia40-4pHA or Fzo1pHA as in (C).