Visualization of eukaryotic DNA mismatch repair reveals distinct recognition and repair intermediates

Cell. 2011 Nov 23;147(5):1040-53. doi: 10.1016/j.cell.2011.10.025.

Abstract

DNA mismatch repair (MMR) increases replication fidelity by eliminating mispaired bases resulting from replication errors. In Saccharomyces cerevisiae, mispairs are primarily detected by the Msh2-Msh6 complex and corrected following recruitment of the Mlh1-Pms1 complex. Here, we visualized functional fluorescent versions of Msh2-Msh6 and Mlh1-Pms1 in living cells. We found that the Msh2-Msh6 complex is an S phase component of replication centers independent of mispaired bases; this localized pool accounted for 10%-15% of MMR in wild-type cells but was essential for MMR in the absence of Exo1. Unexpectedly, Mlh1-Pms1 formed nuclear foci that, although dependent on Msh2-Msh6 for formation, rarely colocalized with Msh2-Msh6 replication-associated foci. Mlh1-Pms1 foci increased when the number of mispaired bases was increased; in contrast, Msh2-Msh6 foci were unaffected. These findings suggest the presence of replication machinery-coupled and -independent pathways for mispair recognition by Msh2-Msh6, which direct formation of superstoichiometric Mlh1-Pms1 foci that represent sites of active MMR.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Mismatch Repair*
  • DNA Repair Enzymes / metabolism
  • DNA Replication
  • DNA-Binding Proteins / metabolism
  • Exodeoxyribonucleases / metabolism
  • Humans
  • MutS Homolog 2 Protein / metabolism
  • Proliferating Cell Nuclear Antigen / metabolism
  • Saccharomyces cerevisiae / metabolism

Substances

  • DNA-Binding Proteins
  • G-T mismatch-binding protein
  • Proliferating Cell Nuclear Antigen
  • Exodeoxyribonucleases
  • exodeoxyribonuclease I
  • MutS Homolog 2 Protein
  • DNA Repair Enzymes