Home LiteratureArticle Details
PMID: 19202191 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Human CtIP mediates cell cycle control of DNA end resection and double strand break repair.

The Journal of biological chemistry ·Vol. 284 ·No. 14 ·2009-04-03 ·Pages 9558-65

Huertas P, Jackson SP

Abstract

In G(0) and G(1), DNA double strand breaks are repaired by nonhomologous end joining, whereas in S and G(2), they are also repaired by homologous recombination. The human CtIP protein controls double strand break (DSB) resection, an event that occurs effectively only in S/G(2) and that promotes homologous recombination but not non-homologous end joining. Here, we mutate a highly conserved cyclin-dependent kinase (CDK) target motif in CtIP and reveal that mutating Thr-847 to Ala impairs resection, whereas mutating it to Glu to mimic constitutive phosphorylation does not. Moreover, we show that unlike cells expressing wild-type CtIP, cells expressing the Thr-to-Glu mutant resect DSBs even after CDK inhibition. Finally, we establish that Thr-847 mutations to either Ala or Glu affect DSB repair efficiency, cause hypersensitivity toward DSB-generating agents, and affect the frequency and nature of radiation-induced chromosomal rearrangements. These results suggest that CDK-mediated control of resection in human cells operates by mechanisms similar to those recently established in yeast.

MeSH Terms
Amino Acid Sequence Animals Carrier Proteins/chemistry,genetics,metabolism Cell Cycle/drug effects Cell Line, Tumor Conserved Sequence Cyclin-Dependent Kinases/antagonists & inhibitors,metabolism DNA/genetics,metabolism DNA Damage/genetics DNA Repair/genetics Endodeoxyribonucleases Genomic Instability Humans Molecular Sequence Data Mutation/genetics Nuclear Proteins/chemistry,genetics,metabolism Protein Kinase Inhibitors/pharmacology Sequence Alignment
Chemicals
Carrier Proteins Nuclear Proteins Protein Kinase Inhibitors DNA Cyclin-Dependent Kinases Endodeoxyribonucleases RBBP8 protein, human
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Huertas Pablo
Gurdon Institute and Department of Zoology, University of Cambridge, Cambridge CB2 1QN, United Kingdom.
Jackson Stephen P
References (35)
35 references, click to expand
  1. Regulation of DNA double-strand break repair pathway choice.
    Cell Res. 2008 Jan;18(1):134-47 PMID: 18157161
  2. Preferential localization of hyperphosphorylated replication protein A to double-strand break repair and checkpoint complexes upon DNA damage.
    Biochem J. 2005 Nov 1;391(Pt 3):473-80 PMID: 15929725
  3. Human CtIP promotes DNA end resection.
    Nature. 2007 Nov 22;450(7169):509-14 PMID: 17965729
  4. Molecular characterization of the role of the Schizosaccharomyces pombe nip1+/ctp1+ gene in DNA double-strand break repair in association with the Mre11-Rad50-Nbs1 complex.
    Mol Cell Biol. 2008 Jun;28(11):3639-51 PMID: 18378696
  5. Molecular views of recombination proteins and their control.
    Nat Rev Mol Cell Biol. 2003 Jun;4(6):435-45 PMID: 12778123
  6. MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks.
    Cell. 2005 Dec 29;123(7):1213-26 PMID: 16377563
  7. Replication-mediated DNA damage by camptothecin induces phosphorylation of RPA by DNA-dependent protein kinase and dissociates RPA:DNA-PK complexes.
    EMBO J. 1999 Mar 1;18(5):1397-406 PMID: 10064605
  8. CDK targets Sae2 to control DNA-end resection and homologous recombination.
    Nature. 2008 Oct 2;455(7213):689-92 PMID: 18716619
  9. Characterization of a carboxy-terminal BRCA1 interacting protein.
    Oncogene. 1998 Nov 5;17(18):2279-85 PMID: 9811458
  10. Screening for microsatellite instability target genes in colorectal cancers.
    J Med Genet. 2002 Nov;39(11):785-9 PMID: 12414815
  11. Differential usage of non-homologous end-joining and homologous recombination in double strand break repair.
    DNA Repair (Amst). 2006 Sep 8;5(9-10):1021-9 PMID: 16807135
  12. Replication protein A phosphorylation and the cellular response to DNA damage.
    DNA Repair (Amst). 2004 Aug-Sep;3(8-9):1015-24 PMID: 15279788
  13. The mechanism of human nonhomologous DNA end joining.
    J Biol Chem. 2008 Jan 4;283(1):1-5 PMID: 17999957
  14. Phospho-dependent interactions between NBS1 and MDC1 mediate chromatin retention of the MRN complex at sites of DNA damage.
    EMBO Rep. 2008 Aug;9(8):795-801 PMID: 18583988
  15. Molecular cloning and characterization of a novel retinoblastoma-binding protein.
    Genomics. 1998 Aug 1;51(3):351-8 PMID: 9721205
  16. Sae2 is an endonuclease that processes hairpin DNA cooperatively with the Mre11/Rad50/Xrs2 complex.
    Mol Cell. 2007 Nov 30;28(4):638-51 PMID: 18042458
  17. Inactivation of CtIP leads to early embryonic lethality mediated by G1 restraint and to tumorigenesis by haploid insufficiency.
    Mol Cell Biol. 2005 May;25(9):3535-42 PMID: 15831459
  18. ATM- and cell cycle-dependent regulation of ATR in response to DNA double-strand breaks.
    Nat Cell Biol. 2006 Jan;8(1):37-45 PMID: 16327781
  19. Analysis of the defect in DNA end joining in the murine scid mutation.
    Mol Cell Biol. 1992 Oct;12(10):4758-68 PMID: 1406659
  20. The functions of budding yeast Sae2 in the DNA damage response require Mec1- and Tel1-dependent phosphorylation.
    Mol Cell Biol. 2004 May;24(10):4151-65 PMID: 15121837
  21. The CDK regulates repair of double-strand breaks by homologous recombination during the cell cycle.
    EMBO J. 2004 Dec 8;23(24):4868-75 PMID: 15549137
  22. Enhanced DNA-PK-mediated RPA2 hyperphosphorylation in DNA polymerase eta-deficient human cells treated with cisplatin and oxaliplatin.
    DNA Repair (Amst). 2008 Apr 2;7(4):582-96 PMID: 18289945
  23. CtIP, a multivalent adaptor connecting transcriptional regulation, checkpoint control and tumor suppression.
    Cell Cycle. 2006 Aug;5(15):1592-6 PMID: 16880746
  24. Ctp1 is a cell-cycle-regulated protein that functions with Mre11 complex to control double-strand break repair by homologous recombination.
    Mol Cell. 2007 Oct 12;28(1):134-46 PMID: 17936710
  25. Differential involvement of phosphatidylinositol 3-kinase-related protein kinases in hyperphosphorylation of replication protein A2 in response to replication-mediated DNA double-strand breaks.
    Genes Cells. 2006 Mar;11(3):237-46 PMID: 16483312
  26. Choreography of the DNA damage response: spatiotemporal relationships among checkpoint and repair proteins.
    Cell. 2004 Sep 17;118(6):699-713 PMID: 15369670
  27. CtIP activates its own and cyclin D1 promoters via the E2F/RB pathway during G1/S progression.
    Mol Cell Biol. 2006 Apr;26(8):3124-34 PMID: 16581787
  28. DNA end resection, homologous recombination and DNA damage checkpoint activation require CDK1.
    Nature. 2004 Oct 21;431(7011):1011-7 PMID: 15496928
  29. A novel plant gene essential for meiosis is related to the human CtIP and the yeast COM1/SAE2 gene.
    EMBO J. 2007 Dec 12;26(24):5061-70 PMID: 18007598
  30. Functional interactions between Sae2 and the Mre11 complex.
    Genetics. 2008 Feb;178(2):711-23 PMID: 18245357
  31. The C-terminal (BRCT) domains of BRCA1 interact in vivo with CtIP, a protein implicated in the CtBP pathway of transcriptional repression.
    J Biol Chem. 1998 Sep 25;273(39):25388-92 PMID: 9738006
  32. Cell cycle-dependent complex formation of BRCA1.CtIP.MRN is important for DNA double-strand break repair.
    J Biol Chem. 2008 Mar 21;283(12):7713-20 PMID: 18171670
  33. TRB3 interacts with CtIP and is overexpressed in certain cancers.
    Biochim Biophys Acta. 2007 Feb;1770(2):273-8 PMID: 17112672
  34. A conserved function for a Caenorhabditis elegans Com1/Sae2/CtIP protein homolog in meiotic recombination.
    EMBO J. 2007 Dec 12;26(24):5071-82 PMID: 18007596
  35. DNA damage-induced cell cycle checkpoint control requires CtIP, a phosphorylation-dependent binding partner of BRCA1 C-terminal domains.
    Mol Cell Biol. 2004 Nov;24(21):9478-86 PMID: 15485915
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
Published
2009-04-03
Epub
2009-00-07
Pages
9558-65
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2666608
Subset
IM
Grants
Biotechnology and Biological Sciences Research Council · BB/F001665/1 · United Kingdom
Biotechnology and Biological Sciences Research Council · BBF0016651 · United Kingdom
Cancer Research UK · United Kingdom
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]