Reconstitution of Long and Short Patch Mismatch Repair Reactions Using Saccharomyces Cerevisiae Proteins
Reconstitution of long and short patch mismatch repair reactions using Saccharomyces cerevisiae proteins Nikki Bowena, Catherine E. Smitha, Anjana Srivatsana, Smaranda Willcoxb,c, Jack D. Griffithb,c, and Richard D. Kolodnera,d,e,f,g,1 aLudwig Institute for Cancer Research, Departments of dMedicine and eCellular and Molecular Medicine, fMoores-University of California, San Diego Cancer Center, and gInstitute of Genomic Medicine, University of California San Diego School of Medicine, La Jolla, CA 92093; and bLineberger Cancer Center and cDepartment of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27514 Contributed by Richard D. Kolodner, October 8, 2013 (sent for review September 9, 2013) A problem in understanding eukaryotic DNA mismatch repair In eukaryotic MMR, mispairs are bound by MutS homolog 2 (MMR) mechanisms is linking insights into MMR mechanisms from (Msh2)–MutS homolog 6 (Msh6) and Msh2–MutS homolog 3 genetics and cell-biology studies with those from biochemical (Msh3), two partially redundant complexes of MutS-related pro- studies of MMR proteins and reconstituted MMR reactions. This teins (3, 4, 18, 19). These complexes recruit a MutL-related type of analysis has proven difficult because reconstitution ap- complex, called MutL homoloh 1 (Mlh1)–postmeiotic segrega- proaches have been most successful for human MMR whereas tion 1 (Pms1) in S. cerevisiae and Mlh1–postmeiotic segregation – – analysis of MMR in vivo has been most advanced in the yeast 2 (Pms2) in human and mouse (3, 4, 20 23). The Mlh1 Pms1/ Saccharomyces cerevisiae. Here, we describe the reconstitution of Pms2 complex has an endonuclease activity suggested to play MMR reactions using purified S.
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