Skip to main content
. 2010 Apr 13;29(10):1713–1725. doi: 10.1038/emboj.2010.61

Figure 3.

Figure 3

The SE domains from BLM/Sgs1 orthologs show strand annealing activity. (A) SA assays contained the indicated concentrations of GST- (upper) or His6-tagged (lower) proteins plus 1 nM each of a 32P-labelled 50 nt oligo (#1) and an unlabelled 50 nt oligo (#2) that share 25 nt of perfect complementarity. The reactions were incubated at 37°C for 5 min under standard conditions as described in ‘Materials and methods'. Reactions were stopped and the products were resolved by 10% PAGE followed by phosphorimager analysis. M is a mock reaction lacking protein. (B) GST–Sgs11−322 (50 nM) or Sgs1103−322 (20 nM) were assayed as in (A) except that the reactions were stopped at the indicated times before analysis. (C) The indicated SE domain proteins (50 nM) were assayed as in (A) except that the reactions contained either no competitor (−) or a 10-, 100- or 1000-fold excess of oligo #16 before the addition of proteins. (D) The indicated SE domain proteins (50 nM) were assayed as in (A) except that the reaction contained 1 mM Mg2+ and either no additions (−) or 1 mM of the indicated cofactor. Throughout, all proteins are His6-tagged unless indicated as GST-tagged. Asterisks represent positions of 32P-labelling.