Extended Data Fig. 3: The scc2-pip mutant shows additive cohesion defects with replisome mutants of the cohesin conversion, but not de novo loading pathway.
From: PCNA recruits cohesin loader Scc2 to ensure sister chromatid cohesion
a, Protein levels of C-terminally 6HA-tagged Scc2 and its PIP mutant variant in elg1Δ wpl1Δ cells. PCNA (yeast Pol30) served as loading control. b-d, Tetrad dissection analysis of the indicated diploid strains. C-terminal 6HA-tagging of Scc2 results in the slow-growth phenotype in elg1Δ wpl1Δ pds5Δ (b), and lethality in elg1Δ wpl1Δ chl1Δ (d) background. e, scc2-pip mutant combined with elg1Δ wpl1Δ pds5Δ results in slow-growth phenotype and strong sensitivity to benomyl. Strains with disruption of the SCC2 terminator by selection marker (tSCC2::HPHNT1) without mutating the SCC2 PIP served as controls. f-h, The scc2-pip mutant, while on its own not sensitive to microtubule poison (f), shows strong sensitivity to benomyl when combined with replisome mutants of the cohesin conversion pathway ctf4Δ, csm3Δ, and tof1Δ (g), but not with the de novo cohesin loading pathway mutant mrc1Δ (h).
