Figure 2.

RSC Recruits the Cohesin Loader Independently of Its ATPase

(A) Sth1^K501R is present on chromatin. Sth1 ChIP was performed in mitotic arrest following Sth1 depletion, as in Figure 1. Binding to three gene promoters (RPL23B, RPL34A, and PUG1) and a tRNA gene (tH(GUG)E1) was measured by real-time qPCR, normalized to a negative control site (CIN8). Means and SEM of three independent experiments are shown. Sth1^Degron + Sth1 wt and Sth1^Degron + Sth1^K501R, p not significant; untagged Sth1, p < 0.01; two-way ANOVA test compared with the endogenous Sth1 strain.

(B) The Sth1 ATPase is required for chromatin remodeling. Shown are average nucleosome occupancy profiles at 4,264 genes aligned to the +1 nucleosome midpoint, comparing wild-type with Sth1^Degron cells following Sth1 depletion and with Sth1^Degron cells expressing either Sth1 or Sth1^K501R.

(C) Cohesin loader recruitment by Sth1^K501R. Scc2 levels were measured by ChIP, followed by real-time qPCR as in (A). Means and SEM of three independent experiments are shown. Sth1^Degron, p < 0.01; Sth1^Degron + Sth1 wild-type and Sth1^Degron + Sth1^K501R, p not significant; two-way ANOVA test compared with the wild-type strain.