title: In vitro studies of DNA mismatch repair proteins.
abstract: The ability to monitor and characterize DNA mismatch repair activity in various mammalian cells is important for understanding mechanisms involved in mutagenesis and tumorigenesis. Since mismatch repair proteins recognize mismatches containing both normal and chemically altered or damaged bases, in vitro assays must accommodate a variety of mismatches in different sequence contexts. Here we describe the construction of DNA mismatch substrates containing G:T or O(6)meG:T mismatches, the purification of recombinant native human MutSα (MSH2-MSH6) and MutLα (MLH1-PMS2) proteins, and in vitro mismatch repair and excision assays that can be adapted to study mismatch repair in nuclear extracts from mismatch repair proficient and deficient cells.
authors.affiliation: Genetics & Biochemistry Branch, National Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Geng
authors.forename: Chunwei
authors.initials: C
authors.lastname: Du
authors.forename: Siying
authors.initials: S
authors.lastname: Chen
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Salerno
authors.forename: Candela
authors.initials: C
authors.lastname: Manfredi
authors.forename: Peggy
authors.initials: P
authors.lastname: Hsieh
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D053843:DNA Mismatch Repair; D045643:DNA Repair Enzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D000070956:MutL Proteins; D011119:Polynucleotides / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C094443:G-T mismatch-binding protein; C500929:MutLalpha protein, human; D011119:Polynucleotides; D011994:Recombinant Proteins; D000070956:MutL Proteins; D045643:DNA Repair Enzymes