FIG. 5.
Pulse-chase analysis of the half-lives of Cdc28 and Cak1 in wild-type and cdc37-1 cells. (A) Wild-type (WT; RD270-8B) and cdc37-1 (RD249-2D p86-1) strains expressing HA epitope-tagged Cdc28 (Cdc28HA) were pulsed with [35S]methionine at 23 or 37°C for 15 min and chased at the same temperature with unlabeled medium for the indicated times. Similar studies were performed with wild-type (WT; AF42) and cdc37-1 (AF40) strains expressing HA epitope-tagged Cak1. Cell lysates were subjected to immunoprecipitation with an anti-HA monoclonal antibody. (B) Wild-type (WT; RD270-8B) and cdc37-1 (RD249-2D p86-1) strains expressing HA epitope-tagged Cdc28 were pulsed with [35S]methionine at 23°C for 30 min and chased with unlabeled medium at 37°C for the indicated times. Similar studies were performed with wild-type (WT; AF42) and cdc37-1 (AF40) strains expressing HA epitope-tagged Cak1. Cell lysates were subjected to immunoprecipitation with an anti-HA monoclonal antibody.