Figure 3.

Rfc1-RFC Promotes DNA Replication but Not Sister Chromatid Cohesion

(A) Cells were synchronized in G1, and Rfc1 was depleted for 2 h by auxin treatment before release into synchronous progression through S phase. DNA replication was monitored by FACS analysis of DNA content.

(B) Cells were synchronized in G1 and released into BrdU-containing medium. Cells were harvested at the indicated times, and BrdU immunoprecipitates were hybridized to Affymetrix GeneChip S. cerevisiae Tiling 1.0R arrays. Signal intensities, relative to a whole-genome DNA sample, normalized to the median BrdU peak intensities, are shown along chromosome 8. Origin positions are indicated.

(C) Boxplots of BrdU peak widths derived from (B) from 52 early origins at the indicated time points.

(D) Cells were arrested in G1, and Rfc1 was depleted for 2 h by auxin treatment before release into HU-containing medium for an early S phase arrest. FLAG-PCNA chromatin immunoprecipitates were analyzed by qPCR with primer pairs around ARS607. Means ± SE from four independent experiments are shown.

(E) Cells were synchronized in G1 and released into nocodazole-containing medium. Sister chromatid cohesion was assessed at the GFP-marked URA3 locus. Means ± SE from three independent experiments are shown.

(F) As in (E), but Smc3 acetylation was quantified relative to total Smc3 levels. Means ± SE from three independent experiments are shown.

See Figure S4 for further characterization of the rfc1-aid strain.