FIG. 2.

Optimal Cbp80p recruitment to GAL1 requires Kin28p-dependent Ser5 phosphorylation of the Pol II CTD. Isogenic CBP80-myc strains CMY013 (KIN28) and CMY014 (kin28-ts16) were cultured in synthetic complete medium lacking histidine with 2% raffinose at 25°C to an A₆₀₀ of ≈0.6 and transferred to 37°C for 30 min, and galactose (2%) was added for another 30 min at 37°C (open bars). The same strains were cultured identically except at 25°C rather than 37°C (filled bars). (A to F) ChIP analysis of factor occupancies at GAL1 was conducted as described for Fig. 1E except with the use of H14 antibodies specific for the Rpb1p CTD phosphorylated on Ser5 (Ser5P) (A and B), Rpb3p antibodies (C and D), or Myc antibodies (to detect Myc-Cbp80p) (E and F). (G) Occupancies of Myc-Cbp80p in panel F were normalized to those for Rpb3p in panel D. The average results obtained from two independent cultures and two PCR amplifications for each culture were plotted in the histograms with standard errors shown as error bars.