Tags

Type your tag names separated by a space and hit enter

Dynamic regulatory interactions of rad51, rad52, and replication protein-a in recombination intermediates.
J Mol Biol. 2009 Jul 03; 390(1):45-55.JM

Abstract

Rad51, Rad52, and replication protein-A (RPA) play crucial roles in the repair of DNA double-strand breaks in Saccharomyces cerevisiae. Rad51 mediates DNA strand exchange, a key reaction in DNA recombination. Rad52 recruits Rad51 into single-stranded DNAs (ssDNAs) that are saturated with RPA. Rad52 also promotes annealing of ssDNA strands that are complexed with RPA. Specific protein-protein interactions are involved in these reactions. Here we report new biochemical characteristics of these protein interactions. First, Rad52-RPA interaction requires multiple molecules of RPA to be associated with ssDNA, suggesting that multiple contacts between the Rad52 ring and RPA-ssDNA filament are needed for stable binding. Second, RPA-t11, which is a recombination-deficient mutant of RPA, displays a defect in interacting with Rad52 in the presence of salt above 50 mM, explaining the defect in Rad52-mediated ssDNA annealing in the presence of this mutation. Third, ssDNA annealing promoted by Rad52 is preceded by aggregation of multiple RPA-ssDNA complexes with Rad52, and Rad51 inhibits this aggregation. These results suggest a regulatory role for Rad51 that suppresses ssDNA annealing and facilitates DNA strand invasion. Finally, the Rad51-double-stranded DNA complex disrupts Rad52-RPA interaction in ssDNA and titrates Rad52 from RPA. This suggests an additional regulatory role for Rad51 following DNA strand invasion, where Rad51-double-stranded DNA may inhibit illegitimate second-end capture to ensure the error-free repair of a DNA double-strand break.

Authors+Show Affiliations

Department of Biological Sciences, Ohio University, Athens, 45701, USA. sugiyama@ohio.eduNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

19445949

Citation

Sugiyama, Tomohiko, and Noriko Kantake. "Dynamic Regulatory Interactions of Rad51, Rad52, and Replication Protein-a in Recombination Intermediates." Journal of Molecular Biology, vol. 390, no. 1, 2009, pp. 45-55.
Sugiyama T, Kantake N. Dynamic regulatory interactions of rad51, rad52, and replication protein-a in recombination intermediates. J Mol Biol. 2009;390(1):45-55.
Sugiyama, T., & Kantake, N. (2009). Dynamic regulatory interactions of rad51, rad52, and replication protein-a in recombination intermediates. Journal of Molecular Biology, 390(1), 45-55. https://doi.org/10.1016/j.jmb.2009.05.009
Sugiyama T, Kantake N. Dynamic Regulatory Interactions of Rad51, Rad52, and Replication Protein-a in Recombination Intermediates. J Mol Biol. 2009 Jul 3;390(1):45-55. PubMed PMID: 19445949.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Dynamic regulatory interactions of rad51, rad52, and replication protein-a in recombination intermediates. AU - Sugiyama,Tomohiko, AU - Kantake,Noriko, Y1 - 2009/05/13/ PY - 2009/02/16/received PY - 2009/05/07/revised PY - 2009/05/07/accepted PY - 2009/5/19/entrez PY - 2009/5/19/pubmed PY - 2009/7/8/medline SP - 45 EP - 55 JF - Journal of molecular biology JO - J Mol Biol VL - 390 IS - 1 N2 - Rad51, Rad52, and replication protein-A (RPA) play crucial roles in the repair of DNA double-strand breaks in Saccharomyces cerevisiae. Rad51 mediates DNA strand exchange, a key reaction in DNA recombination. Rad52 recruits Rad51 into single-stranded DNAs (ssDNAs) that are saturated with RPA. Rad52 also promotes annealing of ssDNA strands that are complexed with RPA. Specific protein-protein interactions are involved in these reactions. Here we report new biochemical characteristics of these protein interactions. First, Rad52-RPA interaction requires multiple molecules of RPA to be associated with ssDNA, suggesting that multiple contacts between the Rad52 ring and RPA-ssDNA filament are needed for stable binding. Second, RPA-t11, which is a recombination-deficient mutant of RPA, displays a defect in interacting with Rad52 in the presence of salt above 50 mM, explaining the defect in Rad52-mediated ssDNA annealing in the presence of this mutation. Third, ssDNA annealing promoted by Rad52 is preceded by aggregation of multiple RPA-ssDNA complexes with Rad52, and Rad51 inhibits this aggregation. These results suggest a regulatory role for Rad51 that suppresses ssDNA annealing and facilitates DNA strand invasion. Finally, the Rad51-double-stranded DNA complex disrupts Rad52-RPA interaction in ssDNA and titrates Rad52 from RPA. This suggests an additional regulatory role for Rad51 following DNA strand invasion, where Rad51-double-stranded DNA may inhibit illegitimate second-end capture to ensure the error-free repair of a DNA double-strand break. SN - 1089-8638 UR - https://www.unboundmedicine.com/medline/citation/19445949/Dynamic_regulatory_interactions_of_rad51_rad52_and_replication_protein_a_in_recombination_intermediates_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(09)00573-7 DB - PRIME DP - Unbound Medicine ER -