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Heteroduplex rejection during single-strand annealing requires Sgs1 helicase and mismatch repair proteins Msh2 and Msh6 but not Pms1.
Proc Natl Acad Sci U S A. 2004 Jun 22; 101(25):9315-20.PN

Abstract

Recombination between moderately divergent DNA sequences is impaired compared with identical sequences. In yeast, an HO endonuclease-induced double-strand break can be repaired by single-strand annealing (SSA) between flanking homologous sequences. A 3% sequence divergence between 205-bp sequences flanking the double-strand break caused a 6-fold reduction in repair compared with identical sequences. This reduction in heteroduplex rejection was suppressed in a mismatch repair-defective msh6 Delta strain and partially suppressed in an msh2 separation-of-function mutant. In mlh1 Delta strains, heteroduplex rejection was greater than in msh6 Delta strains but less than in wild type. Deleting PMS1, MLH2,or MLH3 had no effect on heteroduplex rejection, but a pms1 Delta mlh2 Delta mlh3 Delta triple mutant resembled mlh1 Delta. However, correction of the mismatches within heteroduplex SSA intermediates required PMS1 and MLH1 to the same extent as MSH2 and MSH6. An SSA competition assay in which either diverged or identical repeats can be used for repair showed that heteroduplex DNA is likely to be unwound rather than degraded. This conclusion is supported by the finding that deleting the SGS1 helicase also suppressed heteroduplex rejection.

Authors+Show Affiliations

Rosenstiel Center and Department of Biology, Brandeis University, Waltham, MA 02454-9110, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15199178

Citation

Sugawara, Neal, et al. "Heteroduplex Rejection During Single-strand Annealing Requires Sgs1 Helicase and Mismatch Repair Proteins Msh2 and Msh6 but Not Pms1." Proceedings of the National Academy of Sciences of the United States of America, vol. 101, no. 25, 2004, pp. 9315-20.
Sugawara N, Goldfarb T, Studamire B, et al. Heteroduplex rejection during single-strand annealing requires Sgs1 helicase and mismatch repair proteins Msh2 and Msh6 but not Pms1. Proc Natl Acad Sci U S A. 2004;101(25):9315-20.
Sugawara, N., Goldfarb, T., Studamire, B., Alani, E., & Haber, J. E. (2004). Heteroduplex rejection during single-strand annealing requires Sgs1 helicase and mismatch repair proteins Msh2 and Msh6 but not Pms1. Proceedings of the National Academy of Sciences of the United States of America, 101(25), 9315-20.
Sugawara N, et al. Heteroduplex Rejection During Single-strand Annealing Requires Sgs1 Helicase and Mismatch Repair Proteins Msh2 and Msh6 but Not Pms1. Proc Natl Acad Sci U S A. 2004 Jun 22;101(25):9315-20. PubMed PMID: 15199178.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Heteroduplex rejection during single-strand annealing requires Sgs1 helicase and mismatch repair proteins Msh2 and Msh6 but not Pms1. AU - Sugawara,Neal, AU - Goldfarb,Tamara, AU - Studamire,Barbara, AU - Alani,Eric, AU - Haber,James E, Y1 - 2004/06/15/ PY - 2004/6/17/pubmed PY - 2004/8/24/medline PY - 2004/6/17/entrez SP - 9315 EP - 20 JF - Proceedings of the National Academy of Sciences of the United States of America JO - Proc Natl Acad Sci U S A VL - 101 IS - 25 N2 - Recombination between moderately divergent DNA sequences is impaired compared with identical sequences. In yeast, an HO endonuclease-induced double-strand break can be repaired by single-strand annealing (SSA) between flanking homologous sequences. A 3% sequence divergence between 205-bp sequences flanking the double-strand break caused a 6-fold reduction in repair compared with identical sequences. This reduction in heteroduplex rejection was suppressed in a mismatch repair-defective msh6 Delta strain and partially suppressed in an msh2 separation-of-function mutant. In mlh1 Delta strains, heteroduplex rejection was greater than in msh6 Delta strains but less than in wild type. Deleting PMS1, MLH2,or MLH3 had no effect on heteroduplex rejection, but a pms1 Delta mlh2 Delta mlh3 Delta triple mutant resembled mlh1 Delta. However, correction of the mismatches within heteroduplex SSA intermediates required PMS1 and MLH1 to the same extent as MSH2 and MSH6. An SSA competition assay in which either diverged or identical repeats can be used for repair showed that heteroduplex DNA is likely to be unwound rather than degraded. This conclusion is supported by the finding that deleting the SGS1 helicase also suppressed heteroduplex rejection. SN - 0027-8424 UR - https://www.unboundmedicine.com/medline/citation/15199178/Heteroduplex_rejection_during_single_strand_annealing_requires_Sgs1_helicase_and_mismatch_repair_proteins_Msh2_and_Msh6_but_not_Pms1_ DB - PRIME DP - Unbound Medicine ER -