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. 2016 Jun 3;26(7):775–786. doi: 10.1038/cr.2016.66

Figure 2.

Figure 2

MutSβ stimulates Polβ-induced (CAG)n and (CTG)n hairpin retention during DNA synthesis. Unless mentioned otherwise, hairpin retention/removal assays were performed in a 40-μL purified system containing 0.15 pmol (CAG)5 or (CTG)5 DNA hairpin substrate, 4 pmol MutSβ, 110 fmol RFC and 2 pmol PCNA in addition to the indicated polymerase (600 fmol Polδ, 260 fmol Polβ). Primer extension products were analyzed by Southern blot analysis as described in Figure 1 legends. (A) Effect of MutSβ on Polβ- and Polδ-catalyzed DNA synthesis using a (CAG)5 hairpin as primer. (B) Increased hairpin retention activity of Polβ is proportional to the increasing concerntrations of MutSβ. (C) Effect of MutSβ on Polβ- and Polδ-catalyzed DNA synthesis using a (CTG)5 hairpin as primer. (D) Effect of MutSα:MutSβ ratio on Polβ-catalyzed DNA synthesis using a (CAG)5 hairpin as primer. Relative hairpin retention activity in each group was calculated by using the hairpin retention activity conducted by Polβ alone as a reference, i.e., dividing hairpin retention level of each reaction with that catalyzed by Polβ alone (see *). The data in A-D were from three independent experiments and the error bar represents SD. (E) Proposed DNA structures for shortened DNA products.