FIGURE 6:

Rlm1 physically interacts with SWI/SNF. (A) Rlm1 associates directly with the SWI/SNF complex in vitro. GST, GST-Rlm1, and GST-Rlm1ΔN were expressed and purified from E. coli, and the SWI/SNF complex was purified from a yeast strain bearing TAP-tag Snf2. SWI/SNF complex was incubated with GST proteins bound to glutathione–Sepharose beads, and interacting proteins were probed using antibodies against GST and Snf2. Specific GST fusion proteins are labeled with an asterisk. Total represents 20% of total input protein. (B) In vivo binding of Rlm1 and SWI/SNF. An rlm1Δ snf2Δ strain was transformed with episomal plasmids expressing Rlm1-Myc and Snf2-HA under their native promoters. Samples were taken from yeast cultures growing in the presence or absence of CR (30 min). Snf2-HA was immunoprecipitated by monoclonal antibodies against HA. Snf2-HA and Rlm1-Myc proteins were detected by Western blotting against HA and Myc epitopes, respectively. Total extract represents 10% of the total input protein.