Analysis of the excision step in human DNA mismatch repair.
The reaction responsible for replication error correction by mismatch repair proceeds via several steps: mismatch recognition, mismatch-provoked excision, repair DNA synthesis, and ligation. Key steps in this process are the recognition and subsequent exonucleolytic removal of the mispair. A minimal system comprised of human MutSalpha (MSH2*MSH6), MutLalpha (MLH1*PMS2), exonuclease I (EXOI), replication protein A (RPA), proliferating cell nuclear antigen (PCNA), and replication factor C (RFC) is sufficient to support mismatch-provoked excision in vitro. This chapter describes methods for analysis of the reconstituted excision reaction.
Genschel
Jochen
J
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina, USA.
Modrich
Paul
P
D000818:Animals; D020137:Base Pair Mismatch; D004249:DNA Damage; D053843:DNA Mismatch Repair; D045643:DNA Repair Enzymes; D004268:DNA-Binding Proteins / Q000378:metabolism; D005090:Exodeoxyribonucleases / Q000378:metabolism; D006801:Humans; D000070956:MutL Proteins; D051718:MutS Homolog 2 Protein / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051716:Replication Protein A / Q000378:metabolism; D051818:Replication Protein C / Q000378:metabolism
D004268:DNA-Binding Proteins; C094443:G-T mismatch-binding protein; C500929:MutLalpha protein, human; D009363:Neoplasm Proteins; D018809:Proliferating Cell Nuclear Antigen; D051716:Replication Protein A; D005090:Exodeoxyribonucleases; C028450:exodeoxyribonuclease I; C497172:MSH2 protein, human; D000070956:MutL Proteins; D051718:MutS Homolog 2 Protein; D051818:Replication Protein C; D045643:DNA Repair Enzymes