Biochemical Properties of MutL, a DNA Mismatch Repair Endonuclease

DNA mismatch repair (MMR) is one of the most widely conserved DNA repair systems, which repairs mismatched bases generated mainly by the error of DNA polymerases during replication (Friedberg, et al., 2006, Iyer, et al., 2006, Kunkel, et al., 2005, Morita, et al., 2010). MMR increases the replication fidelity by 20 to 400-fold (Schaaper, 1993). Mutations and epigenetic silencing in MMR genes cause human hereditary nonpolyposis colon cancers as well as sporadic tumors (Fishel, et al., 1995, Fishel, et al., 1994, Kane, et al., 1997, Leach, et al., 1993, Modrich, et al., 1996, Suter, et al., 2004), indicating the significance of this repair system. To date, two types of MMR mechanisms have been clarified: one is employed by eukaryotes and most bacteria (Fig. 1A and B) (Modrich, 2006) and the other is specific to Escherichia coli and other -proteobacteria (Fig. 1C) (Modrich, et al., 1996). The fundamental mechanism and the required proteins in the two types of MMRs are relatively similar to each other. A mismatch is recognized by the bacterial MutS homodimer, eukaryotic MutS (MSH2-MSH6 heterodimer), or MutS (MSH2-MSH3 heterodimer) (Acharya, et al., 2003, Drotschmann, et al., 2002, Gradia, et al., 1997, Gradia, et al., 1999, Lamers, et al., 2000, McCulloch, et al., 2003, Obmolova, et al., 2000, Tachiki, et al., 2000). Subsequently, the bacterial MutL homodimer or eukaryotic MutL (MLH1-PMS2 and MLH1-PMS1 heterodimers in humans and yeast, respectively) is recruited to the mismatched DNA to stimulate downstream events (Acharya, et al., 2003, Kadyrov, et al., 2006). The largest difference between the two types of MMR mechanisms is in the “strand discrimination” system. Although both bases constituting the mismatch are canonical, MMR needs to identify which base is to be repaired. In eukaryotes and most bacteria, MMR directs the repair to the error-containing strand of the mismatched duplex by recognizing the strand discontinuities in the newly synthesized strand (Kadyrov, et al., 2006, Kadyrov, et al., 2007, Larrea, et al., 2010, Modrich, 2006). The termini of leading and lagging strands are thought to serve as discrimination signals. On the other hand, E. coli MMR reads the absence of methylation at the restriction site in the newly synthesized strand (Iyer, et al., 2006, Kunkel, et al., 2005, Larrea, et al., 2010). Before the site-specific DNA methylase (e.g., E. coli Dam methylase (Schlagman, et