Fig. 2.

Sse1 physically interacts with Slt2 and is required for its activation. a FLAG immunoprecipitations were performed on protein extracts derived from W303 cells harboring p416GPD-FLAG-SLT2 or p416GPD as described in “Materials and methods.” Immunoblot analysis using anti-Sse1 and anti-FLAG antibodies was carried out on the extracts and immunoprecipitates. The extract lanes represent 5% of the total amount of extract used for the immunoprecipitation. b A strain carrying an integrated 13×-myc-tagged SLT2 allele (wild type, BY2207) and the isogenic SSE1 deletion (sse1Δ, LSY19) were grown to mid-log phase at 25°C followed by shifting of the cultures to 37°C and removal of aliquots at 0, 30, and 60 min postshift. Protein extracts were prepared followed by anti-myc and anti-phospho-Slt2 immunoblot analysis. c Wild type (DS10), sse1Δ (KMY69), swi4Δ (LSY1), and rlm1Δ (LSY2) cells were transformed with the PST1-lacZ reporter plasmid to measure transcriptional activation by Rlm1 and β-galactosidase activity assayed before and after a 1.5 h heat shock at 37°C