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DNA length dependence of the single-strand annealing pathway and the role of Saccharomyces cerevisiae RAD59 in double-strand break repair.
Mol Cell Biol. 2000 Jul; 20(14):5300-9.MC

Abstract

A DNA double-strand break (DSB) created by the HO endonuclease in Saccharomyces cerevisiae will stimulate recombination between flanking repeats by the single-strand annealing (SSA) pathway, producing a deletion. Previously the efficiency of SSA, using homologous sequences of different lengths, was measured in competition with that of a larger repeat further from the DSB, which ensured that nearly all cells would survive the DSB if the smaller region was not used (N. Sugawara and J. E. Haber, Mol. Cell. Biol. 12:563-575, 1992). Without competition, the efficiency with which homologous segments of 63 to 205 bp engaged in SSA was significantly increased. A sequence as small as 29 bp was used 0.2% of the time, and homology dependence was approximately linear up to 415 bp, at which size almost all cells survived. A mutant with a deletion of RAD59, a homologue of RAD52, was defective for SSA, especially when the homologous-sequence length was short; however, even with 1.17-kb substrates, SSA was reduced fourfold. DSB-induced gene conversion also showed a partial dependence on Rad59p, again being greatest when the homologous-sequence length was short. We found that Rad59p plays a role in removing nonhomologous sequences from the ends of single-stranded DNA when it invades a homologous DNA template, in a manner similar to that previously seen with srs2 mutants. Deltarad59 affected DSB-induced gene conversion differently from msh3 and msh2, which are also defective in removing nonhomologous ends in both DSB-induced gene conversion and SSA. A msh3 rad59 double mutant was more severely defective in SSA than either single mutant.

Authors+Show Affiliations

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

Pub Type(s)

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

Language

eng

PubMed ID

10866686

Citation

Sugawara, N, et al. "DNA Length Dependence of the Single-strand Annealing Pathway and the Role of Saccharomyces Cerevisiae RAD59 in Double-strand Break Repair." Molecular and Cellular Biology, vol. 20, no. 14, 2000, pp. 5300-9.
Sugawara N, Ira G, Haber JE. DNA length dependence of the single-strand annealing pathway and the role of Saccharomyces cerevisiae RAD59 in double-strand break repair. Mol Cell Biol. 2000;20(14):5300-9.
Sugawara, N., Ira, G., & Haber, J. E. (2000). DNA length dependence of the single-strand annealing pathway and the role of Saccharomyces cerevisiae RAD59 in double-strand break repair. Molecular and Cellular Biology, 20(14), 5300-9.
Sugawara N, Ira G, Haber JE. DNA Length Dependence of the Single-strand Annealing Pathway and the Role of Saccharomyces Cerevisiae RAD59 in Double-strand Break Repair. Mol Cell Biol. 2000;20(14):5300-9. PubMed PMID: 10866686.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - DNA length dependence of the single-strand annealing pathway and the role of Saccharomyces cerevisiae RAD59 in double-strand break repair. AU - Sugawara,N, AU - Ira,G, AU - Haber,J E, PY - 2000/6/24/pubmed PY - 2000/8/1/medline PY - 2000/6/24/entrez SP - 5300 EP - 9 JF - Molecular and cellular biology JO - Mol Cell Biol VL - 20 IS - 14 N2 - A DNA double-strand break (DSB) created by the HO endonuclease in Saccharomyces cerevisiae will stimulate recombination between flanking repeats by the single-strand annealing (SSA) pathway, producing a deletion. Previously the efficiency of SSA, using homologous sequences of different lengths, was measured in competition with that of a larger repeat further from the DSB, which ensured that nearly all cells would survive the DSB if the smaller region was not used (N. Sugawara and J. E. Haber, Mol. Cell. Biol. 12:563-575, 1992). Without competition, the efficiency with which homologous segments of 63 to 205 bp engaged in SSA was significantly increased. A sequence as small as 29 bp was used 0.2% of the time, and homology dependence was approximately linear up to 415 bp, at which size almost all cells survived. A mutant with a deletion of RAD59, a homologue of RAD52, was defective for SSA, especially when the homologous-sequence length was short; however, even with 1.17-kb substrates, SSA was reduced fourfold. DSB-induced gene conversion also showed a partial dependence on Rad59p, again being greatest when the homologous-sequence length was short. We found that Rad59p plays a role in removing nonhomologous sequences from the ends of single-stranded DNA when it invades a homologous DNA template, in a manner similar to that previously seen with srs2 mutants. Deltarad59 affected DSB-induced gene conversion differently from msh3 and msh2, which are also defective in removing nonhomologous ends in both DSB-induced gene conversion and SSA. A msh3 rad59 double mutant was more severely defective in SSA than either single mutant. SN - 0270-7306 UR - https://www.unboundmedicine.com/medline/citation/10866686/DNA_length_dependence_of_the_single_strand_annealing_pathway_and_the_role_of_Saccharomyces_cerevisiae_RAD59_in_double_strand_break_repair_ L2 - https://journals.asm.org/doi/10.1128/MCB.20.14.5300-5309.2000?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -