Home LiteratureArticle Details
PMID: 11459964 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Rad52 forms DNA repair and recombination centers during S phase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 98 ·No. 15 ·2001-07-17 ·Pages 8276-82

Lisby M, Rothstein R, Mortensen UH

Abstract

Maintenance of genomic integrity and stable transmission of genetic information depend on a number of DNA repair processes. Failure to faithfully perform these processes can result in genetic alterations and subsequent development of cancer and other genetic diseases. In the eukaryote Saccharomyces cerevisiae, homologous recombination is the major pathway for repairing DNA double-strand breaks. The key role played by Rad52 in this pathway has been attributed to its ability to seek out and mediate annealing of homologous DNA strands. In this study, we find that S. cerevisiae Rad52 fused to green fluorescent protein (GFP) is fully functional in DNA repair and recombination. After induction of DNA double-strand breaks by gamma-irradiation, meiosis, or the HO endonuclease, Rad52-GFP relocalizes from a diffuse nuclear distribution to distinct foci. Interestingly, Rad52 foci are formed almost exclusively during the S phase of mitotic cells, consistent with coordination between recombinational repair and DNA replication. This notion is further strengthened by the dramatic increase in the frequency of Rad52 focus formation observed in a pol12-100 replication mutant and a mec1 DNA damage checkpoint mutant. Furthermore, our data indicate that each Rad52 focus represents a center of recombinational repair capable of processing multiple DNA lesions.

MeSH Terms
Cell Nucleus/metabolism DNA Damage DNA Repair DNA, Fungal/biosynthesis DNA-Binding Proteins/genetics,metabolism,physiology Deoxyribonucleases, Type II Site-Specific/metabolism Fungal Proteins/genetics,metabolism,physiology Intracellular Signaling Peptides and Proteins Mitosis Protein Serine-Threonine Kinases Rad52 DNA Repair and Recombination Protein Recombinant Fusion Proteins/genetics,metabolism,physiology Recombination, Genetic S Phase Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Signal Transduction
Chemicals
DNA, Fungal DNA-Binding Proteins Fungal Proteins Intracellular Signaling Peptides and Proteins RAD52 protein, S cerevisiae Rad52 DNA Repair and Recombination Protein Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins MEC1 protein, S cerevisiae Protein Serine-Threonine Kinases HO protein, S cerevisiae SCEI protein, S cerevisiae Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lisby M
Department of Genetics and Development, Columbia University, College of Physicians and Surgeons, 701 West 168th Street, New York, NY 10032-2704, USA.
Rothstein R
Mortensen U H
References (65)
65 references, click to expand
  1. 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
  2. Mating-type gene switching in Saccharomyces cerevisiae.
    Trends Genet. 1992 Dec;8(12):446-52 PMID: 1492369
  3. DNA double-strand breaks and the RAD50-RAD57 genes in Saccharomyces.
    Semin Cancer Biol. 1993 Apr;4(2):73-83 PMID: 8513150
  4. 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
  5. Activation of a yeast replication origin near a double-stranded DNA break.
    Genes Dev. 1994 Mar 1;8(5):554-62 PMID: 7926750
  6. 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
  7. 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
  8. Radiation mutagenesis: the initial DNA lesions responsible.
    Radiat Res. 1995 Jun;142(3):362-8 PMID: 7761586
  9. Thermosensitivity of green fluorescent protein fluorescence utilized to reveal novel nuclear-like compartments in a mutant nucleoporin NSP1.
    J Biochem. 1995 Jul;118(1):13-7 PMID: 8537302
  10. Specific interactions between the human RAD51 and RAD52 proteins.
    J Biol Chem. 1996 Jan 5;271(1):148-52 PMID: 8550550
  11. Gene conversion plays the major role in controlling the stability of large tandem repeats in yeast.
    EMBO J. 1996 Apr 1;15(7):1715-25 PMID: 8612596
  12. Crystal structure of the Aequorea victoria green fluorescent protein.
    Science. 1996 Sep 6;273(5280):1392-5 PMID: 8703075
  13. Organization of DNA into foci during replication.
    Curr Opin Cell Biol. 1996 Jun;8(3):365-8 PMID: 8743888
  14. A Rad52 homolog is required for RAD51-independent mitotic recombination in Saccharomyces cerevisiae.
    Genes Dev. 1996 Aug 15;10(16):2025-37 PMID: 8769646
  15. 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
  16. A novel mutation avoidance mechanism dependent on S. cerevisiae RAD27 is distinct from DNA mismatch repair.
    Cell. 1997 Jan 24;88(2):253-63 PMID: 9008166
  17. When checkpoints fail.
    Cell. 1997 Feb 7;88(3):315-21 PMID: 9039258
  18. Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family.
    Cell. 1997 Feb 7;88(3):375-84 PMID: 9039264
  19. Identification of novel isoforms of human RAD52.
    Biochim Biophys Acta. 1999 Dec 23;1489(2-3):303-14 PMID: 10673031
  20. The human Rad52 protein exists as a heptameric ring.
    Curr Biol. 2000 Mar 23;10(6):337-40 PMID: 10744977
  21. 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
  22. Recombination factors of Saccharomyces cerevisiae.
    Mutat Res. 2000 Jun 30;451(1-2):257-75 PMID: 10915877
  23. Precise binding of single-stranded DNA termini by human RAD52 protein.
    EMBO J. 2000 Aug 1;19(15):4175-81 PMID: 10921897
  24. Rad22 protein, a rad52 homologue in Schizosaccharomyces pombe, binds to DNA double-strand breaks.
    J Biol Chem. 2000 Nov 10;275(45):35607-11 PMID: 10956666
  25. Characterization of RAD52 homologs in the fission yeast Schizosaccharomyces pombe.
    Mutat Res. 2001 Jan 5;461(4):311-23 PMID: 11104907
  26. Coordinated response of mammalian Rad51 and Rad52 to DNA damage.
    EMBO Rep. 2000 Jul;1(1):85-90 PMID: 11256631
  27. The repair of double-strand breaks in the nuclear DNA of Saccharomyces cerevisiae and its genetic control.
    Mol Gen Genet. 1976 Jan 16;143(2):119-29 PMID: 765749
  28. Saccharomyces cerevisiae pol30 (proliferating cell nuclear antigen) mutations impair replication fidelity and mismatch repair.
    Mol Cell Biol. 1999 Nov;19(11):7801-15 PMID: 10523669
  29. The double-strand-break repair model for recombination.
    Cell. 1983 May;33(1):25-35 PMID: 6380756
  30. Directionality and regulation of cassette substitution in yeast.
    Cold Spring Harb Symp Quant Biol. 1984;49:97-104 PMID: 6397325
  31. Analysis of meiosis-defective mutations in yeast by physical monitoring of recombination.
    Genetics. 1986 Jul;113(3):551-67 PMID: 3015718
  32. Homothallic mating type switching generates lethal chromosome breaks in rad52 strains of Saccharomyces cerevisiae.
    Mol Cell Biol. 1981 Jun;1(6):522-34 PMID: 6765605
  33. Elevated recombination rates in transcriptionally active DNA.
    Cell. 1989 Feb 24;56(4):619-30 PMID: 2645056
  34. Two DNA repair and recombination genes in Saccharomyces cerevisiae, RAD52 and RAD54, are induced during meiosis.
    Mol Cell Biol. 1989 Jul;9(7):3101-4 PMID: 2506437
  35. A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae.
    Cell. 1990 Jun 15;61(6):1089-101 PMID: 2190690
  36. Multifunctional yeast high-copy-number shuttle vectors.
    Gene. 1992 Jan 2;110(1):119-22 PMID: 1544568
  37. 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
  38. A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae.
    Cell. 1983 Nov;35(1):167-74 PMID: 6313222
  39. Holliday junctions accumulate in replication mutants via a RecA homolog-independent mechanism.
    Cell. 1997 Jul 11;90(1):87-96 PMID: 9230305
  40. Replication protein A: a heterotrimeric, single-stranded DNA-binding protein required for eukaryotic DNA metabolism.
    Annu Rev Biochem. 1997;66:61-92 PMID: 9242902
  41. Meiotic chromosomes: it takes two to tango.
    Genes Dev. 1997 Oct 15;11(20):2600-21 PMID: 9334324
  42. Function of yeast Rad52 protein as a mediator between replication protein A and the Rad51 recombinase.
    J Biol Chem. 1997 Nov 7;272(45):28194-7 PMID: 9353267
  43. The complete set of predicted genes from Saccharomyces cerevisiae in a readily usable form.
    Genome Res. 1997 Dec;7(12):1169-73 PMID: 9414322
  44. Cloning-free PCR-based allele replacement methods.
    Genome Res. 1997 Dec;7(12):1174-83 PMID: 9414323
  45. Stimulation by Rad52 of yeast Rad51-mediated recombination.
    Nature. 1998 Jan 22;391(6665):404-7 PMID: 9450759
  46. Rad52 protein stimulates DNA strand exchange by Rad51 and replication protein A.
    Nature. 1998 Jan 22;391(6665):407-10 PMID: 9450760
  47. DNA annealing by RAD52 protein is stimulated by specific interaction with the complex of replication protein A and single-stranded DNA.
    Proc Natl Acad Sci U S A. 1998 May 26;95(11):6049-54 PMID: 9600915
  48. Recombination at work for meiosis.
    Curr Opin Genet Dev. 1998 Apr;8(2):200-11 PMID: 9610411
  49. Rad52 forms ring structures and co-operates with RPA in single-strand DNA annealing.
    Genes Cells. 1998 Mar;3(3):145-56 PMID: 9619627
  50. 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
  51. Rad52 associates with RPA and functions with rad55 and rad57 to assemble meiotic recombination complexes.
    Genes Dev. 1998 Jul 15;12(14):2208-21 PMID: 9679065
  52. Chromosomal rearrangements occur in S. cerevisiae rfa1 mutator mutants due to mutagenic lesions processed by double-strand-break repair.
    Mol Cell. 1998 Jul;2(1):9-22 PMID: 9702187
  53. DNA damage and checkpoint pathways: molecular anatomy and interactions with repair.
    Cell. 1998 Sep 4;94(5):555-8 PMID: 9741620
  54. A suppressor of two essential checkpoint genes identifies a novel protein that negatively affects dNTP pools.
    Mol Cell. 1998 Sep;2(3):329-40 PMID: 9774971
  55. 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
  56. 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
  57. Visualisation of human rad52 protein and its complexes with hRad51 and DNA.
    J Mol Biol. 1998 Dec 11;284(4):1027-38 PMID: 9837724
  58. Homologous recombination is required for the viability of rad27 mutants.
    Nucleic Acids Res. 1998 Dec 15;26(24):5589-95 PMID: 9837987
  59. An allele of RFA1 suppresses RAD52-dependent double-strand break repair in Saccharomyces cerevisiae.
    Genetics. 1999 Feb;151(2):447-58 PMID: 9927442
  60. Double-strand break repair in yeast requires both leading and lagging strand DNA polymerases.
    Cell. 1999 Feb 5;96(3):415-24 PMID: 10025407
  61. Binding of double-strand breaks in DNA by human Rad52 protein.
    Nature. 1999 Apr 22;398(6729):728-31 PMID: 10227297
  62. RAD50 and RAD51 define two pathways that collaborate to maintain telomeres in the absence of telomerase.
    Genetics. 1999 May;152(1):143-52 PMID: 10224249
  63. Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae.
    Microbiol Mol Biol Rev. 1999 Jun;63(2):349-404 PMID: 10357855
  64. Localization and dynamic relocalization of mammalian Rad52 during the cell cycle and in response to DNA damage.
    Curr Biol. 1999 Sep 9;9(17):975-8 PMID: 10508584
  65. A requirement for recombinational repair in Saccharomyces cerevisiae is caused by DNA replication defects of mec1 mutants.
    Genetics. 1999 Oct;153(2):595-605 PMID: 10511542
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.121006298
Published
2001-07-17
Pages
8276-82
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC37432
Subset
IM
Grants
NHGRI NIH HHS · R01 HG001620 · United States
NIGMS NIH HHS · R37 GM050237 · United States
NIGMS NIH HHS · R01 GM050237 · United States
NHGRI NIH HHS · HG01620 · United States
NCRR NIH HHS · S10 RR10506 · United States
NCI NIH HHS · P30 CA013696 · United States
NIGMS NIH HHS · GM50237 · United States
NCI NIH HHS · P30 CA13696 · United States
External Links
PubMed source DOI: 10.1073/pnas.121006298 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2