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PMID: 12724407 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The mechanism of Mus81-Mms4 cleavage site selection distinguishes it from the homologous endonuclease Rad1-Rad10.

Molecular and cellular biology ·Vol. 23 ·No. 10 ·2003-05-00 ·Pages 3487-96

Bastin-Shanower SA, Fricke WM, Mullen JR, Brill SJ

Abstract

Mus81-Mms4 and Rad1-Rad10 are homologous structure-specific endonucleases that cleave 3' branches from distinct substrates and are required for replication fork stability and nucleotide excision repair, respectively, in the yeast Saccharomyces cerevisiae. We explored the basis of this biochemical and genetic specificity. The Mus81-Mms4 cleavage site, a nick 5 nucleotides (nt) 5' of the flap, is determined not by the branch point, like Rad1-Rad10, but by the 5' end of the DNA strand at the flap junction. As a result, the endonucleases show inverse substrate specificity; substrates lacking a 5' end within 4 nt of the flap are cleaved poorly by Mus81-Mms4 but are cleaved well by Rad1-10. Genetically, we show that both mus81 and sgs1 mutants are sensitive to camptothecin-induced DNA damage. Further, mus81 sgs1 synthetic lethality requires homologous recombination, as does suppression of mutant phenotypes by RusA expression. These data are most easily explained by a model in which the in vivo substrate of Mus81-Mms4 and Sgs1-Top3 is a 3' flap recombination intermediate downstream of replication fork collapse.

MeSH Terms
Base Sequence DNA Damage DNA Repair Enzymes DNA-Binding Proteins/metabolism Dimerization Dose-Response Relationship, Drug Dose-Response Relationship, Radiation Endonucleases/metabolism Flap Endonucleases Fungal Proteins/metabolism Molecular Sequence Data Mutation Plasmids/metabolism Protein Binding Recombinant Proteins/metabolism Recombination, Genetic Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins Single-Strand Specific DNA and RNA Endonucleases Substrate Specificity Time Factors Trans-Activators/metabolism Ultraviolet Rays
Chemicals
DNA-Binding Proteins Fungal Proteins Recombinant Proteins Saccharomyces cerevisiae Proteins Trans-Activators Endonucleases Flap Endonucleases MUS81 protein, S cerevisiae RAD1 protein, S cerevisiae MMS4 protein, S cerevisiae RAD10 protein, S cerevisiae Single-Strand Specific DNA and RNA Endonucleases DNA Repair Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bastin-Shanower Suzanne A
Department of Molecular Biology and Biochemistry, CABM, Rutgers University, 679 Hoes Lane, Piscataway, NJ 08854, USA.
Fricke William M
Mullen Janet R
Brill Steven J
References (48)
48 references, click to expand
  1. Replication fork pausing and recombination or "gimme a break".
    Genes Dev. 2000 Jan 1;14(1):1-10 PMID: 10640269
  2. Systematic genetic analysis with ordered arrays of yeast deletion mutants.
    Science. 2001 Dec 14;294(5550):2364-8 PMID: 11743205
  3. The Bloom's syndrome gene product interacts with topoisomerase III.
    J Biol Chem. 2000 Mar 31;275(13):9636-44 PMID: 10734115
  4. Bipartite structure of the SGS1 DNA helicase in Saccharomyces cerevisiae.
    Genetics. 2000 Mar;154(3):1101-14 PMID: 10757756
  5. 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
  6. MUS81 encodes a novel helix-hairpin-helix protein involved in the response to UV- and methylation-induced DNA damage in Saccharomyces cerevisiae.
    Mol Gen Genet. 2000 Jun;263(5):812-27 PMID: 10905349
  7. Interaction between yeast sgs1 helicase and DNA topoisomerase III.
    J Biol Chem. 2000 Sep 1;275(35):26898-905 PMID: 10862619
  8. Damage tolerance protein Mus81 associates with the FHA1 domain of checkpoint kinase Cds1.
    Mol Cell Biol. 2000 Dec;20(23):8758-66 PMID: 11073977
  9. Requirement for three novel protein complexes in the absence of the Sgs1 DNA helicase in Saccharomyces cerevisiae.
    Genetics. 2001 Jan;157(1):103-18 PMID: 11139495
  10. 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
  11. Mapping the DNA topoisomerase III binding domain of the Sgs1 DNA helicase.
    J Biol Chem. 2001 Mar 23;276(12):8848-55 PMID: 11124263
  12. Human Mus81-associated endonuclease cleaves Holliday junctions in vitro.
    Mol Cell. 2001 Nov;8(5):1117-27 PMID: 11741546
  13. A role for MMS4 in the processing of recombination intermediates during meiosis in Saccharomyces cerevisiae.
    Genetics. 2001 Dec;159(4):1511-25 PMID: 11779793
  14. Substrate specificity of RusA resolvase reveals the DNA structures targeted by RuvAB and RecG in vivo.
    Mol Cell. 2002 Jul;10(1):187-98 PMID: 12150918
  15. Mus81-Eme1 and Rqh1 involvement in processing stalled and collapsed replication forks.
    J Biol Chem. 2002 Sep 6;277(36):32753-9 PMID: 12084712
  16. Holliday junction resolution in human cells: two junction endonucleases with distinct substrate specificities.
    EMBO J. 2002 Oct 15;21(20):5577-85 PMID: 12374758
  17. Yeast Tdp1 and Rad1-Rad10 function as redundant pathways for repairing Top1 replicative damage.
    Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13669-74 PMID: 12368472
  18. Repair of topoisomerase I covalent complexes in the absence of the tyrosyl-DNA phosphodiesterase Tdp1.
    Proc Natl Acad Sci U S A. 2002 Nov 12;99(23):14970-5 PMID: 12397185
  19. 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
  20. A model for replication repair in mammalian cells.
    J Mol Biol. 1976 Mar 5;101(3):417-25 PMID: 1255724
  21. DNA topoisomerase-targeting antitumor drugs can be studied in yeast.
    Proc Natl Acad Sci U S A. 1988 Oct;85(20):7501-5 PMID: 2845409
  22. Evidence that DNA topoisomerase I is necessary for the cytotoxic effects of camptothecin.
    Mol Pharmacol. 1988 Dec;34(6):755-60 PMID: 2849043
  23. DNA topoisomerases: why so many?
    J Biol Chem. 1991 Apr 15;266(11):6659-62 PMID: 1849888
  24. Identification of the yeast TOP3 gene product as a single strand-specific DNA topoisomerase.
    J Biol Chem. 1992 Aug 25;267(24):17178-85 PMID: 1324925
  25. Sister chromatids are preferred over homologs as substrates for recombinational repair in Saccharomyces cerevisiae.
    Genetics. 1992 Oct;132(2):387-402 PMID: 1427035
  26. Replication-dependent sister chromatid recombination in rad1 mutants of Saccharomyces cerevisiae.
    Genetics. 1993 Mar;133(3):469-87 PMID: 8454200
  27. Yeast DNA repair and recombination proteins Rad1 and Rad10 constitute a single-stranded-DNA endonuclease.
    Nature. 1993 Apr 29;362(6423):860-2 PMID: 8479526
  28. Specific cleavage of model recombination and repair intermediates by the yeast Rad1-Rad10 DNA endonuclease.
    Science. 1994 Sep 30;265(5181):2082-5 PMID: 8091230
  29. The yeast type I topoisomerase Top3 interacts with Sgs1, a DNA helicase homolog: a potential eukaryotic reverse gyrase.
    Mol Cell Biol. 1994 Dec;14(12):8391-8 PMID: 7969174
  30. The structure-selectivity and sequence-preference of the junction-resolving enzyme CCE1 of Saccharomyces cerevisiae.
    J Mol Biol. 1996 Mar 29;257(2):330-41 PMID: 8609627
  31. A new efficient gene disruption cassette for repeated use in budding yeast.
    Nucleic Acids Res. 1996 Jul 1;24(13):2519-24 PMID: 8692690
  32. Xeroderma pigmentosum group F caused by a defect in a structure-specific DNA repair endonuclease.
    Cell. 1996 Sep 6;86(5):811-22 PMID: 8797827
  33. Mechanisms of cell killing by drugs that trap covalent complexes between DNA topoisomerases and DNA.
    Mol Pharmacol. 1996 Nov;50(5):1095-102 PMID: 8913340
  34. Mechanism of action of camptothecin.
    Ann N Y Acad Sci. 1996 Dec 13;803:44-9 PMID: 8993499
  35. recF and recR are required for the resumption of replication at DNA replication forks in Escherichia coli.
    Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3714-9 PMID: 9108043
  36. Holliday junctions accumulate in replication mutants via a RecA homolog-independent mechanism.
    Cell. 1997 Jul 11;90(1):87-96 PMID: 9230305
  37. DNA structural elements required for ERCC1-XPF endonuclease activity.
    J Biol Chem. 1998 Apr 3;273(14):7835-42 PMID: 9525876
  38. Mms4, a putative transcriptional (co)activator, protects Saccharomyces cerevisiae cells from endogenous and environmental DNA damage.
    Mol Gen Genet. 1998 Apr;257(6):614-23 PMID: 9604884
  39. RuvAB acts at arrested replication forks.
    Cell. 1998 Oct 30;95(3):419-30 PMID: 9814711
  40. The essential role of yeast topoisomerase III in meiosis depends on recombination.
    EMBO J. 1999 Mar 15;18(6):1701-11 PMID: 10075939
  41. Defending genome integrity during DNA replication: a proposed role for RecQ family helicases.
    Bioessays. 1999 Apr;21(4):286-94 PMID: 10377891
  42. Three new dominant drug resistance cassettes for gene disruption in Saccharomyces cerevisiae.
    Yeast. 1999 Oct;15(14):1541-53 PMID: 10514571
  43. Historical overview: searching for replication help in all of the rec places.
    Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8173-80 PMID: 11459950
  44. The DNA replication checkpoint response stabilizes stalled replication forks.
    Nature. 2001 Aug 2;412(6846):557-61 PMID: 11484058
  45. Pathways for repair of topoisomerase I covalent complexes in Saccharomyces cerevisiae.
    Genes Cells. 2001 Aug;6(8):677-87 PMID: 11532027
  46. Functional overlap between Sgs1-Top3 and the Mms4-Mus81 endonuclease.
    Genes Dev. 2001 Oct 15;15(20):2730-40 PMID: 11641278
  47. Mus81-Eme1 are essential components of a Holliday junction resolvase.
    Cell. 2001 Nov 16;107(4):537-48 PMID: 11719193
  48. Association of the Bloom syndrome protein with topoisomerase IIIalpha in somatic and meiotic cells.
    Cancer Res. 2000 Mar 1;60(5):1162-7 PMID: 10728666
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.23.10.3487-3496.2003
Published
2003-05-00
Pages
3487-96
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC164751
Subset
IM
Grants
NIA NIH HHS · R01 AG016637-05 · United States
NIA NIH HHS · AG 16637 · United States
External Links
PubMed source DOI: 10.1128/MCB.23.10.3487-3496.2003 PMC Full Text View journal details
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