Home LiteratureArticle Details
PMID: 17353272 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Mus81-Eme1-dependent and -independent crossovers form in mitotic cells during double-strand break repair in Schizosaccharomyces pombe.

Molecular and cellular biology ·Vol. 27 ·No. 10 ·2007-05-00 ·Pages 3828-38

Hope JC, Cruzata LD, Duvshani A, Mitsumoto J, Maftahi M, Freyer GA

Abstract

During meiosis, double-strand breaks (DSBs) lead to crossovers, thought to arise from the resolution of double Holliday junctions (HJs) by an HJ resolvase. In Schizosaccharomyces pombe, meiotic crossovers are produced primarily through a mechanism requiring the Mus81-Eme1 endonuclease complex. Less is known about the processes that produces crossovers during the repair of DSBs in mitotic cells. We employed an inducible DSB system to determine the role of Rqh1-Top3 and Mus81-Eme1 in mitotic DSB repair and crossover formation in S. pombe. In agreement with the meiotic data, crossovers are suppressed in cells lacking Mus81-Eme1. And relative to the wild type, rqh1Delta cells show a fourfold increase in crossover frequency. This suppression of crossover formation by Rqh1 is dependent on its helicase activity. We found that the synthetic lethality of cells lacking both Rqh1 and Eme1 is suppressed by loss of swi5(+), which allowed us to show that the excess crossovers formed in an rqh1Delta background are independent of Mus81-Eme1. This result suggests that a second process for crossover formation exists in S. pombe and is consistent with our finding that deletion of swi5(+) restored meiotic crossovers in eme1Delta cells. Evidence suggesting that Rqh1 also acts downstream of Swi5 in crossover formation was uncovered in these studies. Our results suggest that during Rhp51-dependent repair of DSBs, Rqh1-Top3 suppresses crossovers in the Rhp57-dependent pathway while Mus81-Eme1 and possibly Rqh1 promote crossovers in the Swi5-dependent pathway.

MeSH Terms
Adenosine Triphosphatases/genetics,metabolism Crossing Over, Genetic DNA Breaks, Double-Stranded DNA Helicases/genetics,metabolism DNA Repair DNA Topoisomerases, Type I/genetics,metabolism DNA-Binding Proteins/genetics,metabolism Electrophoresis, Gel, Pulsed-Field Endonucleases/genetics,metabolism Gene Conversion Mitosis/physiology Rad51 Recombinase/genetics,metabolism Schizosaccharomyces/cytology,genetics,physiology Schizosaccharomyces pombe Proteins/genetics,metabolism
Chemicals
DNA-Binding Proteins MUS81 protein, S pombe RHP51 protein, S pombe Schizosaccharomyces pombe Proteins Swi5 protein, S pombe Rad51 Recombinase Eme1 protein, S pombe Endonucleases Adenosine Triphosphatases Rhp57 protein, S pombe DNA Helicases Rqh1 protein, S pombe DNA Topoisomerases, Type I
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Hope Justin C
Graduate Program in Anatomy and Cell Biology, Columbia University, 722 W. 168th Street, New York, NY 10032, USA.
Cruzata Lissette Delgado
Duvshani Amit
Mitsumoto Jun
Maftahi Mohamed
Freyer Greg A
References (54)
54 references, click to expand
  1. Functional overlap between Sgs1-Top3 and the Mms4-Mus81 endonuclease.
    Genes Dev. 2001 Oct 15;15(20):2730-40 PMID: 11641278
  2. Chromosomal stability and the DNA double-stranded break connection.
    Nat Rev Genet. 2001 Mar;2(3):196-206 PMID: 11256071
  3. Chromosome synapsis defects and sexually dimorphic meiotic progression in mice lacking Spo11.
    Mol Cell. 2000 Nov;6(5):989-98 PMID: 11106739
  4. Elevated incidence of loss of heterozygosity (LOH) in an sgs1 mutant of Saccharomyces cerevisiae: roles of yeast RecQ helicase in suppression of aneuploidy, interchromosomal rearrangement, and the simultaneous incidence of both events during mitotic growth.
    Mutat Res. 2002 Jul 25;504(1-2):157-72 PMID: 12106656
  5. Mus81-Eme1 and Rqh1 involvement in processing stalled and collapsed replication forks.
    J Biol Chem. 2002 Sep 6;277(36):32753-9 PMID: 12084712
  6. The severe slow growth of Deltasrs2 Deltarqh1 in Schizosaccharomyces pombe is suppressed by loss of recombination and checkpoint genes.
    Nucleic Acids Res. 2002 Nov 1;30(21):4781-92 PMID: 12409469
  7. Role of RAD52 epistasis group genes in homologous recombination and double-strand break repair.
    Microbiol Mol Biol Rev. 2002 Dec;66(4):630-70, table of contents PMID: 12456786
  8. Helicase activity is only partially required for Schizosaccharomyces pombe Rqh1p function.
    Yeast. 2002 Dec;19(16):1381-98 PMID: 12478586
  9. Alternate pathways involving Sgs1/Top3, Mus81/ Mms4, and Srs2 prevent formation of toxic recombination intermediates from single-stranded gaps created by DNA replication.
    Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16887-92 PMID: 12475932
  10. Cleavage of model replication forks by fission yeast Mus81-Eme1 and budding yeast Mus81-Mms4.
    J Biol Chem. 2003 Feb 28;278(9):6928-35 PMID: 12473680
  11. RecQ helicases: caretakers of the genome.
    Nat Rev Cancer. 2003 Mar;3(3):169-78 PMID: 12612652
  12. Pathway utilization in response to a site-specific DNA double-strand break in fission yeast.
    EMBO J. 2003 Mar 17;22(6):1419-30 PMID: 12628934
  13. The mechanism of Mus81-Mms4 cleavage site selection distinguishes it from the homologous endonuclease Rad1-Rad10.
    Mol Cell Biol. 2003 May;23(10):3487-96 PMID: 12724407
  14. Identification and characterization of the human mus81-eme1 endonuclease.
    J Biol Chem. 2003 Jul 4;278(27):25172-8 PMID: 12721304
  15. The role of DNA breaks in genomic instability and tumorigenesis.
    Immunol Rev. 2003 Aug;194:77-95 PMID: 12846809
  16. The endogenous Mus81-Eme1 complex resolves Holliday junctions by a nick and counternick mechanism.
    Mol Cell. 2003 Sep;12(3):747-59 PMID: 14527419
  17. Generating crossovers by resolution of nicked Holliday junctions: a role for Mus81-Eme1 in meiosis.
    Mol Cell. 2003 Sep;12(3):761-74 PMID: 14527420
  18. Holliday junctions in the eukaryotic nucleus: resolution in sight?
    Trends Biochem Sci. 2003 Oct;28(10):548-57 PMID: 14559184
  19. Eme1 is involved in DNA damage processing and maintenance of genomic stability in mammalian cells.
    EMBO J. 2003 Nov 17;22(22):6137-47 PMID: 14609959
  20. Srs2 and Sgs1-Top3 suppress crossovers during double-strand break repair in yeast.
    Cell. 2003 Nov 14;115(4):401-11 PMID: 14622595
  21. The Bloom's syndrome helicase suppresses crossing over during homologous recombination.
    Nature. 2003 Dec 18;426(6968):870-4 PMID: 14685245
  22. Two different Swi5-containing protein complexes are involved in mating-type switching and recombination repair in fission yeast.
    Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15770-5 PMID: 14663140
  23. Human diseases deficient in RecQ helicases.
    Biochimie. 2003 Nov;85(11):1185-93 PMID: 14726023
  24. The involvement of Srs2 in post-replication repair and homologous recombination in fission yeast.
    Nucleic Acids Res. 2004;32(4):1480-91 PMID: 14993467
  25. A manyfold increase in sister chromatid exchanges in Bloom's syndrome lymphocytes.
    Proc Natl Acad Sci U S A. 1974 Nov;71(11):4508-12 PMID: 4140506
  26. The double-strand-break repair model for recombination.
    Cell. 1983 May;33(1):25-35 PMID: 6380756
  27. Lethality induced by a single site-specific double-strand break in a dispensable yeast plasmid.
    Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5613-7 PMID: 8516308
  28. Evidences for possible involvement of Rhp51 protein in mitotic events including chromosome segregation.
    Biochem Mol Biol Int. 1995 Oct;37(2):329-37 PMID: 8673016
  29. Double-strand break-induced mitotic intrachromosomal recombination in the fission yeast Schizosaccharomyces pombe.
    Genetics. 1996 Feb;142(2):341-57 PMID: 8852835
  30. Fission yeast rad12+ regulates cell cycle checkpoint control and is homologous to the Bloom's syndrome disease gene.
    Mol Cell Biol. 1998 May;18(5):2721-8 PMID: 9566891
  31. Bloom's and Werner's syndrome genes suppress hyperrecombination in yeast sgs1 mutant: implication for genomic instability in human diseases.
    Proc Natl Acad Sci U S A. 1998 Jul 21;95(15):8733-8 PMID: 9671747
  32. Molecular mechanisms of DNA double strand break repair.
    Trends Cell Biol. 1998 Dec;8(12):483-9 PMID: 9861670
  33. Mammalian MutS homologue 5 is required for chromosome pairing in meiosis.
    Nat Genet. 1999 Jan;21(1):123-7 PMID: 9916805
  34. Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae.
    Microbiol Mol Biol Rev. 1999 Jun;63(2):349-404 PMID: 10357855
  35. Topoisomerase III is essential for accurate nuclear division in Schizosaccharomyces pombe.
    Nucleic Acids Res. 1999 Oct 15;27(20):4050-8 PMID: 10497270
  36. Swi5 acts in meiotic DNA joint molecule formation in Schizosaccharomyces pombe.
    Genetics. 2004 Dec;168(4):1891-8 PMID: 15466419
  37. BLM helicase facilitates Mus81 endonuclease activity in human cells.
    Cancer Res. 2005 Apr 1;65(7):2526-31 PMID: 15805243
  38. A postsynaptic role for Rhp55/57 that is responsible for cell death in Deltarqh1 mutants following replication arrest in Schizosaccharomyces pombe.
    Genetics. 2005 Jun;170(2):519-31 PMID: 15802523
  39. BLAP75/RMI1 promotes the BLM-dependent dissolution of homologous recombination intermediates.
    Proc Natl Acad Sci U S A. 2006 Mar 14;103(11):4068-73 PMID: 16537486
  40. Rqh1 blocks recombination between sister chromatids during double strand break repair, independent of its helicase activity.
    Proc Natl Acad Sci U S A. 2006 Apr 11;103(15):5875-80 PMID: 16595622
  41. Sgs1 regulates gene conversion tract lengths and crossovers independently of its helicase activity.
    Mol Cell Biol. 2006 Jun;26(11):4086-94 PMID: 16705162
  42. Double-strand-break-induced homologous recombination in mammalian cells.
    Biochem Soc Trans. 2001 May;29(Pt 2):196-201 PMID: 11356153
  43. Mechanism and control of meiotic recombination initiation.
    Curr Top Dev Biol. 2001;52:1-53 PMID: 11529427
  44. Single Holliday junctions are intermediates of meiotic recombination.
    Cell. 2006 Dec 15;127(6):1167-78 PMID: 17174892
  45. The Swi5-Sfr1 complex stimulates Rhp51/Rad51- and Dmc1-mediated DNA strand exchange in vitro.
    Nat Struct Mol Biol. 2006 Sep;13(9):823-30 PMID: 16921379
  46. The top3(+) gene is essential in Schizosaccharomyces pombe and the lethality associated with its loss is caused by Rad12 helicase activity.
    Nucleic Acids Res. 1999 Dec 15;27(24):4715-24 PMID: 10572171
  47. A recombination repair gene of Schizosaccharomyces pombe, rhp57, is a functional homolog of the Saccharomyces cerevisiae RAD57 gene and is phylogenetically related to the human XRCC3 gene.
    Genetics. 2000 Apr;154(4):1451-61 PMID: 10747044
  48. Homologous recombination is responsible for cell death in the absence of the Sgs1 and Srs2 helicases.
    Nat Genet. 2000 Jun;25(2):192-4 PMID: 10835635
  49. Partial suppression of the fission yeast rqh1(-) phenotype by expression of a bacterial Holliday junction resolvase.
    EMBO J. 2000 Jun 1;19(11):2751-62 PMID: 10835372
  50. Damage tolerance protein Mus81 associates with the FHA1 domain of checkpoint kinase Cds1.
    Mol Cell Biol. 2000 Dec;20(23):8758-66 PMID: 11073977
  51. SGS1, the Saccharomyces cerevisiae homologue of BLM and WRN, suppresses genome instability and homeologous recombination.
    Nat Genet. 2001 Jan;27(1):113-6 PMID: 11138010
  52. Mutations in recombinational repair and in checkpoint control genes suppress the lethal combination of srs2Delta with other DNA repair genes in Saccharomyces cerevisiae.
    Genetics. 2001 Feb;157(2):557-65 PMID: 11156978
  53. DNA double-strand breaks: signaling, repair and the cancer connection.
    Nat Genet. 2001 Mar;27(3):247-54 PMID: 11242102
  54. Mus81-Eme1 are essential components of a Holliday junction resolvase.
    Cell. 2001 Nov 16;107(4):537-48 PMID: 11719193
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.01596-06
Published
2007-05-00
Epub
2007-00-12
Pages
3828-38
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1900003
Subset
IM
Grants
NIGMS NIH HHS · F31 GM020376 · United States
NCI NIH HHS · R01 CA072647 · United States
NCI NIH HHS · CA 072647 · United States
NIGMS NIH HHS · GM 20376 · United States
External Links
PubMed source DOI: 10.1128/MCB.01596-06 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