Home LiteratureArticle Details
PMID: 12384589 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

DNA end-binding specificity of human Rad50/Mre11 is influenced by ATP.

Nucleic acids research ·Vol. 30 ·No. 20 ·2002-10-15 ·Pages 4425-31

de Jager M, Wyman C, van Gent DC, Kanaar R

Abstract

The Rad50, Mre11 and Nbs1 complex is involved in many essential chromosomal organization processes dealing with DNA ends, including two major pathways of DNA double-strand break repair, homologous recombination and non-homologous end joining. Previous data on the structure of the human Rad50 and Mre11 (R/M) complex suggest that a common role for the protein complex in these processes is to provide a physical link between DNA ends such that they can be processed in an organized and coordinated manner. Here we describe the DNA binding properties of the R/M complex. The complex bound to both single-stranded and double-stranded DNA. Scanning force microscopy analysis of DNA binding by R/M showed the requirement for an end to form oligomeric R/M complexes, which could then migrate or transfer away from the end. The R/M complex had a lower preference for DNA substrates with 3'-overhangs compared with blunt ends or 5'-overhangs. Interestingly, ATP binding, but not hydrolysis, increased the preference of R/M binding to DNA substrates with 3'-overhangs relative to substrates with blunt ends and 5'-overhangs.

MeSH Terms
Acid Anhydride Hydrolases Adenosine Triphosphate/metabolism,pharmacology Animals Cell Line DNA/metabolism,ultrastructure DNA Repair Enzymes DNA, Single-Stranded/metabolism DNA-Binding Proteins/isolation & purification,metabolism Humans MRE11 Homologue Protein Macromolecular Substances Microscopy, Atomic Force Protein Binding
Chemicals
DNA, Single-Stranded DNA-Binding Proteins MRE11 protein, human Macromolecular Substances Adenosine Triphosphate DNA MRE11 Homologue Protein Acid Anhydride Hydrolases Rad50 protein, human DNA Repair Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
de Jager Martijn
Department of Cell Biology and Genetics, Erasmus MC, PO Box 1738, 3000 DR Rotterdam, The Netherlands.
Wyman Claire
van Gent Dik C
Kanaar Roland
References (47)
47 references, click to expand
  1. Ku is associated with the telomere in mammals.
    Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12454-8 PMID: 10535943
  2. Disruption of mRad50 causes embryonic stem cell lethality, abnormal embryonic development, and sensitivity to ionizing radiation.
    Proc Natl Acad Sci U S A. 1999 Jun 22;96(13):7376-81 PMID: 10377422
  3. Replication fork pausing and recombination or "gimme a break".
    Genes Dev. 2000 Jan 1;14(1):1-10 PMID: 10640269
  4. The importance of repairing stalled replication forks.
    Nature. 2000 Mar 2;404(6773):37-41 PMID: 10716434
  5. Initiation of genetic recombination and recombination-dependent replication.
    Trends Biochem Sci. 2000 Apr;25(4):156-65 PMID: 10754547
  6. A mechanistic basis for Mre11-directed DNA joining at microhomologies.
    Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6409-14 PMID: 10823903
  7. Cell-cycle-regulated association of RAD50/MRE11/NBS1 with TRF2 and human telomeres.
    Nat Genet. 2000 Jul;25(3):347-52 PMID: 10888888
  8. Structural biology of Rad50 ATPase: ATP-driven conformational control in DNA double-strand break repair and the ABC-ATPase superfamily.
    Cell. 2000 Jun 23;101(7):789-800 PMID: 10892749
  9. Mre11 and Rad50 from Pyrococcus furiosus: cloning and biochemical characterization reveal an evolutionarily conserved multiprotein machine.
    J Bacteriol. 2000 Nov;182(21):6036-41 PMID: 11029422
  10. Response to RAG-mediated VDJ cleavage by NBS1 and gamma-H2AX.
    Science. 2000 Dec 8;290(5498):1962-5 PMID: 11110662
  11. DNA-binding and strand-annealing activities of human Mre11: implications for its roles in DNA double-strand break repair pathways.
    Nucleic Acids Res. 2001 Mar 15;29(6):1317-25 PMID: 11238998
  12. Targeted disruption of the Nijmegen breakage syndrome gene NBS1 leads to early embryonic lethality in mice.
    Curr Biol. 2001 Jan 23;11(2):105-9 PMID: 11231126
  13. Structural biochemistry and interaction architecture of the DNA double-strand break repair Mre11 nuclease and Rad50-ATPase.
    Cell. 2001 May 18;105(4):473-85 PMID: 11371344
  14. Rescue of arrested replication forks by homologous recombination.
    Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8181-8 PMID: 11459951
  15. The architecture of the human Rad54-DNA complex provides evidence for protein translocation along DNA.
    Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8454-60 PMID: 11459989
  16. Structure of the Ku heterodimer bound to DNA and its implications for double-strand break repair.
    Nature. 2001 Aug 9;412(6847):607-14 PMID: 11493912
  17. DNA structure-specific nuclease activities in the Saccharomyces cerevisiae Rad50*Mre11 complex.
    J Biol Chem. 2001 Sep 21;276(38):35458-64 PMID: 11454871
  18. A Ku bridge over broken DNA.
    Structure. 2001 Oct;9(10):881-4 PMID: 11591342
  19. Human Rad50/Mre11 is a flexible complex that can tether DNA ends.
    Mol Cell. 2001 Nov;8(5):1129-35 PMID: 11741547
  20. Recombination at double-strand breaks and DNA ends: conserved mechanisms from phage to humans.
    Mol Cell. 2001 Dec;8(6):1163-74 PMID: 11779493
  21. DNA double-strand break repair from head to tail.
    Curr Opin Struct Biol. 2002 Feb;12(1):115-22 PMID: 11839498
  22. Protection of mammalian telomeres.
    Oncogene. 2002 Jan 21;21(4):532-40 PMID: 11850778
  23. Targeted disruption of NBS1 reveals its roles in mouse development and DNA repair.
    EMBO J. 2002 Mar 15;21(6):1447-55 PMID: 11889050
  24. The Rad50 zinc-hook is a structure joining Mre11 complexes in DNA recombination and repair.
    Nature. 2002 Aug 1;418(6897):562-6 PMID: 12152085
  25. Genome instability and Rad50(S): subtle yet severe.
    Genes Dev. 2002 Sep 1;16(17):2173-8 PMID: 12208840
  26. Cancer predisposition and hematopoietic failure in Rad50(S/S) mice.
    Genes Dev. 2002 Sep 1;16(17):2237-51 PMID: 12208847
  27. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold.
    EMBO J. 1982;1(8):945-51 PMID: 6329717
  28. HeLa nuclear protein recognizing DNA termini and translocating on DNA forming a regular DNA-multimeric protein complex.
    J Mol Biol. 1989 Jul 5;208(1):65-78 PMID: 2769755
  29. Analysis of the mechanism of interaction of simian Ku protein with DNA.
    Nucleic Acids Res. 1991 Oct 25;19(20):5619-24 PMID: 1945839
  30. RAD50 protein of S.cerevisiae exhibits ATP-dependent DNA binding.
    Nucleic Acids Res. 1993 Aug 11;21(16):3851-6 PMID: 8367302
  31. Purification and characterization of the human Rad51 protein, an analogue of E. coli RecA.
    EMBO J. 1994 Dec 1;13(23):5764-71 PMID: 7988572
  32. Unusual oligomerization required for activity of NtrC, a bacterial enhancer-binding protein.
    Science. 1997 Mar 14;275(5306):1658-61 PMID: 9054362
  33. Conditional gene targeted deletion by Cre recombinase demonstrates the requirement for the double-strand break repair Mre11 protein in murine embryonic stem cells.
    Nucleic Acids Res. 1997 Aug 1;25(15):2985-91 PMID: 9224597
  34. Overexpression, purification, and characterization of the SbcCD protein from Escherichia coli.
    J Biol Chem. 1997 Aug 8;272(32):19819-26 PMID: 9242643
  35. Ku80 is required for immunoglobulin isotype switching.
    EMBO J. 1998 Apr 15;17(8):2404-11 PMID: 9545251
  36. Nibrin, a novel DNA double-strand break repair protein, is mutated in Nijmegen breakage syndrome.
    Cell. 1998 May 1;93(3):467-76 PMID: 9590180
  37. The hMre11/hRad50 protein complex and Nijmegen breakage syndrome: linkage of double-strand break repair to the cellular DNA damage response.
    Cell. 1998 May 1;93(3):477-86 PMID: 9590181
  38. Ku70 is required for late B cell development and immunoglobulin heavy chain class switching.
    J Exp Med. 1998 Jun 15;187(12):2081-9 PMID: 9625768
  39. The 3' to 5' exonuclease activity of Mre 11 facilitates repair of DNA double-strand breaks.
    Mol Cell. 1998 Jun;1(7):969-79 PMID: 9651580
  40. Nuclease activities in a complex of human recombination and DNA repair factors Rad50, Mre11, and p95.
    J Biol Chem. 1998 Aug 21;273(34):21447-50 PMID: 9705271
  41. The human RAD54 recombinational DNA repair protein is a double-stranded DNA-dependent ATPase.
    J Biol Chem. 1998 Oct 23;273(43):28292-7 PMID: 9774452
  42. The many interfaces of Mre11.
    Cell. 1998 Nov 25;95(5):583-6 PMID: 9845359
  43. Molecular mechanisms of DNA double strand break repair.
    Trends Cell Biol. 1998 Dec;8(12):483-9 PMID: 9861670
  44. Conserved domains in DNA repair proteins and evolution of repair systems.
    Nucleic Acids Res. 1999 Mar 1;27(5):1223-42 PMID: 9973609
  45. Nbs1 potentiates ATP-driven DNA unwinding and endonuclease cleavage by the Mre11/Rad50 complex.
    Genes Dev. 1999 May 15;13(10):1276-88 PMID: 10346816
  46. Ku, a DNA repair protein with multiple cellular functions?
    Mutat Res. 1999 May 14;434(1):3-15 PMID: 10377944
  47. The DNA double-strand break repair gene hMRE11 is mutated in individuals with an ataxia-telangiectasia-like disorder.
    Cell. 1999 Dec 10;99(6):577-87 PMID: 10612394
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkf574
Published
2002-10-15
Pages
4425-31
Language
English
Region
England
NLM ID
0411011
PMCID
PMC137138
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkf574 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: product@genelibs.com

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