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PMID: 11606529 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The yeast recombinational repair protein Rad59 interacts with Rad52 and stimulates single-strand annealing.

Genetics ·Vol. 159 ·No. 2 ·2001-10-00 ·Pages 515-25

Davis AP, Symington LS

Abstract

The yeast RAD52 gene is essential for homology-dependent repair of DNA double-strand breaks. In vitro, Rad52 binds to single- and double-stranded DNA and promotes annealing of complementary single-stranded DNA. Genetic studies indicate that the Rad52 and Rad59 proteins act in the same recombination pathway either as a complex or through overlapping functions. Here we demonstrate physical interaction between Rad52 and Rad59 using the yeast two-hybrid system and co-immunoprecipitation from yeast extracts. Purified Rad59 efficiently anneals complementary oligonucleotides and is able to overcome the inhibition to annealing imposed by replication protein A (RPA). Although Rad59 has strand-annealing activity by itself in vitro, this activity is insufficient to promote strand annealing in vivo in the absence of Rad52. The rfa1-D288Y allele partially suppresses the in vivo strand-annealing defect of rad52 mutants, but this is independent of RAD59. These results suggest that in vivo Rad59 is unable to compete with RPA for single-stranded DNA and therefore is unable to promote single-strand annealing. Instead, Rad59 appears to augment the activity of Rad52 in strand annealing.

MeSH Terms
Base Sequence DNA Primers DNA Repair/genetics DNA, Fungal/genetics DNA, Single-Stranded/genetics DNA-Binding Proteins/metabolism Fungal Proteins/metabolism Mutation Precipitin Tests Protein Binding Rad52 DNA Repair and Recombination Protein Recombination, Genetic Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Sequence Deletion
Chemicals
DNA Primers DNA, Fungal DNA, Single-Stranded DNA-Binding Proteins Fungal Proteins RAD52 protein, S cerevisiae RAD59 protein, S cerevisiae Rad52 DNA Repair and Recombination Protein Saccharomyces cerevisiae Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Davis A P
Department of Microbiology and Institute of Cancer Research, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA.
Symington L S
References (52)
52 references, click to expand
  1. A novel allele of RAD52 that causes severe DNA repair and recombination deficiencies only in the absence of RAD51 or RAD59.
    Genetics. 1999 Nov;153(3):1117-30 PMID: 10545446
  2. Double-strand breaks stimulate alternative mechanisms of recombination repair.
    J Mol Biol. 1989 Jun 5;207(3):527-41 PMID: 2668534
  3. The human Rad52 protein exists as a heptameric ring.
    Curr Biol. 2000 Mar 23;10(6):337-40 PMID: 10744977
  4. Functional interactions among yeast Rad51 recombinase, Rad52 mediator, and replication protein A in DNA strand exchange.
    J Biol Chem. 2000 May 26;275(21):15895-904 PMID: 10748203
  5. 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 PMID: 10866686
  6. Galactose as a gratuitous inducer of GAL gene expression in yeasts growing on glucose.
    Gene. 1989 Nov 15;83(1):57-64 PMID: 2512199
  7. Intermediates of recombination during mating type switching in Saccharomyces cerevisiae.
    EMBO J. 1990 Mar;9(3):663-73 PMID: 2178924
  8. Characterization of double-strand break-induced recombination: homology requirements and single-stranded DNA formation.
    Mol Cell Biol. 1992 Feb;12(2):563-75 PMID: 1732731
  9. Rad51 protein involved in repair and recombination in S. cerevisiae is a RecA-like protein.
    Cell. 1992 May 1;69(3):457-70 PMID: 1581961
  10. Dominant negative alleles of RAD52 reveal a DNA repair/recombination complex including Rad51 and Rad52.
    Genes Dev. 1993 Sep;7(9):1755-65 PMID: 8370524
  11. Catalysis of ATP-dependent homologous DNA pairing and strand exchange by yeast RAD51 protein.
    Science. 1994 Aug 26;265(5176):1241-3 PMID: 8066464
  12. Use of a chromosomal inverted repeat to demonstrate that the RAD51 and RAD52 genes of Saccharomyces cerevisiae have different roles in mitotic recombination.
    Genetics. 1994 Nov;138(3):587-95 PMID: 7851757
  13. Protein-protein interactions in the yeast pheromone response pathway: Ste5p interacts with all members of the MAP kinase cascade.
    Genetics. 1994 Nov;138(3):609-19 PMID: 7851759
  14. A novel allele of Saccharomyces cerevisiae RFA1 that is deficient in recombination and repair and suppressible by RAD52.
    Mol Cell Biol. 1995 Mar;15(3):1620-31 PMID: 7862153
  15. A mutation in the gene encoding the Saccharomyces cerevisiae single-stranded DNA-binding protein Rfa1 stimulates a RAD52-independent pathway for direct-repeat recombination.
    Mol Cell Biol. 1995 Mar;15(3):1632-41 PMID: 7862154
  16. Complex formation in yeast double-strand break repair: participation of Rad51, Rad52, Rad55, and Rad57 proteins.
    Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6925-9 PMID: 7624345
  17. Functional differences and interactions among the putative RecA homologs Rad51, Rad55, and Rad57.
    Mol Cell Biol. 1995 Sep;15(9):4843-50 PMID: 7651402
  18. Targeted disruption of the Rad51 gene leads to lethality in embryonic mice.
    Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6236-40 PMID: 8692798
  19. Physical interaction between human RAD52 and RPA is required for homologous recombination in mammalian cells.
    J Biol Chem. 1996 Aug 2;271(31):18996-9000 PMID: 8702565
  20. A Rad52 homolog is required for RAD51-independent mitotic recombination in Saccharomyces cerevisiae.
    Genes Dev. 1996 Aug 15;10(16):2025-37 PMID: 8769646
  21. Genetic requirements for the single-strand annealing pathway of double-strand break repair in Saccharomyces cerevisiae.
    Genetics. 1996 Mar;142(3):693-704 PMID: 8849880
  22. DNA strand annealing is promoted by the yeast Rad52 protein.
    Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10729-34 PMID: 8855248
  23. Direct association between the yeast Rad51 and Rad54 recombination proteins.
    J Biol Chem. 1996 Dec 27;271(52):33181-6 PMID: 8969173
  24. Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast.
    Genetics. 1996 Dec;144(4):1425-36 PMID: 8978031
  25. SIR2 and SIR4 interactions differ in core and extended telomeric heterochromatin in yeast.
    Genes Dev. 1997 Jan 1;11(1):83-93 PMID: 9000052
  26. Yeast Rad55 and Rad57 proteins form a heterodimer that functions with replication protein A to promote DNA strand exchange by Rad51 recombinase.
    Genes Dev. 1997 May 1;11(9):1111-21 PMID: 9159392
  27. Recombinational repair in yeast: functional interactions between Rad51 and Rad54 proteins.
    EMBO J. 1997 May 1;16(9):2535-44 PMID: 9171366
  28. Synergistic actions of Rad51 and Rad52 in recombination and DNA repair.
    Nature. 1998 Jan 22;391(6665):401-4 PMID: 9450758
  29. Stimulation by Rad52 of yeast Rad51-mediated recombination.
    Nature. 1998 Jan 22;391(6665):404-7 PMID: 9450759
  30. Rad52 protein stimulates DNA strand exchange by Rad51 and replication protein A.
    Nature. 1998 Jan 22;391(6665):407-10 PMID: 9450760
  31. Requirement for end-joining and checkpoint functions, but not RAD52-mediated recombination, after EcoRI endonuclease cleavage of Saccharomyces cerevisiae DNA.
    Mol Cell Biol. 1998 Apr;18(4):1891-902 PMID: 9528760
  32. Catalysis of homologous DNA pairing by yeast Rad51 and Rad54 proteins.
    Nature. 1998 May 7;393(6680):91-4 PMID: 9590697
  33. Rad52 forms ring structures and co-operates with RPA in single-strand DNA annealing.
    Genes Cells. 1998 Mar;3(3):145-56 PMID: 9619627
  34. Studies of the interaction between Rad52 protein and the yeast single-stranded DNA binding protein RPA.
    Mol Cell Biol. 1998 Jul;18(7):4400-6 PMID: 9632824
  35. Targeted inactivation of mouse RAD52 reduces homologous recombination but not resistance to ionizing radiation.
    Mol Cell Biol. 1998 Nov;18(11):6423-9 PMID: 9774658
  36. Homologous recombination, but not DNA repair, is reduced in vertebrate cells deficient in RAD52.
    Mol Cell Biol. 1998 Nov;18(11):6430-5 PMID: 9774659
  37. Complex formation and functional versatility of Mre11 of budding yeast in recombination.
    Cell. 1998 Nov 25;95(5):705-16 PMID: 9845372
  38. An allele of RFA1 suppresses RAD52-dependent double-strand break repair in Saccharomyces cerevisiae.
    Genetics. 1999 Feb;151(2):447-58 PMID: 9927442
  39. Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae.
    Microbiol Mol Biol Rev. 1999 Jun;63(2):349-404 PMID: 10357855
  40. Characterization of the role played by the RAD59 gene of Saccharomyces cerevisiae in ectopic recombination.
    Curr Genet. 1999 Aug;36(1-2):13-20 PMID: 10447590
  41. A double-strand break repair component is essential for S phase completion in fission yeast cell cycling.
    Mol Biol Cell. 1999 Oct;10(10):3331-43 PMID: 10512870
  42. Precise binding of single-stranded DNA termini by human RAD52 protein.
    EMBO J. 2000 Aug 1;19(15):4175-81 PMID: 10921897
  43. Superhelicity-driven homologous DNA pairing by yeast recombination factors Rad51 and Rad54.
    Mol Cell. 2000 Sep;6(3):563-72 PMID: 11030336
  44. Rad54 protein is targeted to pairing loci by the Rad51 nucleoprotein filament.
    Mol Cell. 2000 Sep;6(3):583-92 PMID: 11030338
  45. Aberrant double-strand break repair in rad51 mutants of Saccharomyces cerevisiae.
    Mol Cell Biol. 2000 Dec;20(24):9162-72 PMID: 11094068
  46. Characterization of RAD52 homologs in the fission yeast Schizosaccharomyces pombe.
    Mutat Res. 2001 Jan 5;461(4):311-23 PMID: 11104907
  47. Two survivor pathways that allow growth in the absence of telomerase are generated by distinct telomere recombination events.
    Mol Cell Biol. 2001 Mar;21(5):1819-27 PMID: 11238918
  48. Genetic requirements for RAD51- and RAD54-independent break-induced replication repair of a chromosomal double-strand break.
    Mol Cell Biol. 2001 Mar;21(6):2048-56 PMID: 11238940
  49. Isolation and characterization of the RAD59 homologue of Kluyveromyces lactis.
    Curr Genet. 2001 Jul;39(5-6):305-10 PMID: 11525403
  50. A genetic study of x-ray sensitive mutants in yeast.
    Mutat Res. 1974 Sep;24(3):281-92 PMID: 4606119
  51. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics.
    Methods Enzymol. 1987;154:164-75 PMID: 3323810
  52. RAD51 is required for the repair of plasmid double-stranded DNA gaps from either plasmid or chromosomal templates.
    Mol Cell Biol. 2000 Feb;20(4):1194-205 PMID: 10648605
Article Info
Journal
Genetics
Abbr.
Genetics
ISSN
0016-6731
DOI
10.1093/genetics/159.2.515
Published
2001-10-00
Pages
515-25
Language
English
Region
United States
NLM ID
0374636
PMCID
PMC1461847
Subset
IM
Grants
NIGMS NIH HHS · R01 GM041784 · United States
NIGMS NIH HHS · GM41784 · United States
NIAID NIH HHS · T32 AI07161 · United States
NCI NIH HHS · T32 CA09503 · United States
External Links
PubMed source DOI: 10.1093/genetics/159.2.515 PMC Full Text View journal details
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