FIGURE 5.

Identification of novel Slt2 phosphorylation targets. A, Western blot analysis of the in vitro kinase assay carried out as in Fig. 3B but using E. coli cell extracts containing recombinant GST-Rcn2, GST-Gga1, GST-Caf20, or GST-Sod1 as substrate. B, Western blot analysis of the in vitro kinase assay similar to that described in Fig. 3B but using yeast extracts containing GST, GST-Msg5, GST-Mkk1, GST-Rcn2, GST-Gga1, GST-Caf20, or GST-Sod1 as substrate. Anti-thiophosphate (Anti-ThioP) ester antibodies (right panels) and anti-GST (left panels) were used to detect thiophosphorylated proteins and GST-tagged proteins, respectively. Numbers below the blot indicate the amount of thiophosphorylated GST-Msg5, GST-Mkk1, GST-Rcn2, GST-Caf20, and GST-Gga1 in the kinase assay using GST-Slt2-as normalized with respect to their amount in the control kinase assay with GST.