FIG. 4.

Nps1/RSC transiently binds to the IME2 TATA box. (A) Time course of Nps1-TAP binding to the IME2 TATA sequence. The strains used were WHK40-D (IME1) and WMY14-D (ime1Δ). The amount of immunoprecipitated DNA was determined by PCR with primer pairs directed against the IME2 TATA, calculated as described in the legend to Fig. 2B and shown as a relative amount referring to the value of the YPD-grown cells (Y) of IME1 as 1. As a control, a primer set for HTA1 ORF was used. The ratio of the TATA signal to the HTA1 signal of untagged samples was almost equivalent to that of input samples. The values are averages of three independent experiments. (B) Ime1-HA is copurified with Nps1-TAP. The NPS1-TAP IME1-HA (WMY15-D) or NPS1 IME1-HA (minus TAP tag; WMY11-D) cells were harvested at the indicated times and processed for TAP. Proteins eluted from a calmodulin-Sepharose gel were concentrated by lyophilization and then immunoblotted with anti-HA and anti-Nps1 antibodies.