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. Author manuscript; available in PMC: 2016 Feb 25.
Published in final edited form as: Mol Cell. 2010 Sep 10;39(5):689–699. doi: 10.1016/j.molcel.2010.08.008

Figure 6. Acetylation Stabilizes Cohesion.

Figure 6

Strains K17513 (256LacO::TRP1, PMET3CDC20, YEp-PGAL1HOS1, SMC3-PK6), K17514 (256La-cO::TRP1, PMET3CDC20, YEp-PGAL1, SMC3-PK6), K17383 (tetO112::URA3, PMET3CDC20, YEp-PGAL1HOS1), K17384 (tetO112::URA3, MET3-CDC20, YEp-PGAL1), and K9027 (MATα, te-tO112::URA3, PMET3CDC20, scc1Δ, SCC1-TEV268-HA3, GAL-TEV) were grown to log phase in synthetic medium lacking methionine and arrested in metaphase for 2 hr in YEP medium sup-plemented with 2 mM methionine and 2% raffinose (0 min). Expression of Hos1 was induced by addition of 2% galactose (0-180 min).

(A) Proteins from the strains K17513 and K17514 were extracted, and the levels of Smc3 acetylation and Smc3-PK6 at the indicated time points were monitored by western blotting of crude extracts using α-pk and α-acetylated Smc3 antibodies.

(B) Cohesion was measured by scoring the percentages of separated GFP dots at the TRP1 (12.6 kb from CEN4) and URA3 (38.4 kb from CEN5) loci.