Figure 9. The PIP-motif of xMutSα is important for retention of the MMR capability derived from a gap.
(A) Kinetics of the gap-filling reaction in MutS/MutL-depleted NPE containing 630 nM of xMutSαWT, xMutSαPIP, or xMutSαΔN. (B) The efficiencies of the gap-filling reaction were calculated from three independent experiments including the one shown in (A) and plotted onto a graph. Lines: one-phase decay fitting. See Table 2 for the fitting parameters. (C) Kinetics of MMR after supplying fresh NPE to the reaction described in (A). Addition of neither MutS-depleted nor MutL-depleted NPE alone, but both restored MMR in MutS/MutL-depleted NPE (lanes 2–4), showing both MutSα/β and MutLα/β/γ were functionally depleted. (D) The efficiencies of MMR were calculated and presented as described in (B).
Figure 9—figure supplement 1. Depletion efficiencies of Msh2 and Mlh1 from NPE.
NPE was depleted using a mixture of Msh2, Msh6, and Mlh1 antibodies (lanes 1–4), a mixture of Msh2 and Msh6 antibodies (lane 5) or Mlh1 antibodies (lane 6) and supplemented with either control buffer, 630 nM of xMutSαWT, xMutSαPIP, or xMutSαΔN. The depletion efficiencies were estimated to be >98% for both Msh2 and Mlh1.