ATM mediated phosphorylation of CHD4 contributes to genome maintenance

BackgroundIn order to maintain cellular viability and genetic integrity cells must respond quickly following the induction of cytotoxic double strand DNA breaks (DSB). This response requires a number of processes including stabilisation of the DSB, signalling of the break and repair. It is becoming increasingly apparent that one key step in this process is chromatin remodelling.ResultsHere we describe the chromodomain helicase DNA-binding protein (CHD4) as a target of ATM kinase. We show that ionising radiation (IR)-induced phosphorylation of CHD4 affects its intranuclear organization resulting in increased chromatin binding/retention. We also show assembly of phosphorylated CHD4 foci at sites of DNA damage, which might be required to fulfil its function in the regulation of DNA repair. Consistent with this, cells overexpressing a phospho-mutant version of CHD4 that cannot be phosphorylated by ATM fail to show enhanced chromatin retention after DSBs and display high rates of spontaneous damage.ConclusionThese results provide insight into how CHD4 phosphorylation might be required to remodel chromatin around DNA breaks allowing efficient DNA repair to occur.

[1]  R. Gibbons.  Histone modifying and chromatin remodelling enzymes in cancer and dysplastic syndromes. , 2005, Human molecular genetics.

[2]  John A Tainer,et al.  Mre11-Rad50-Nbs1 is a keystone complex connecting DNA repair machinery, double-strand break signaling, and the chromatin template. , 2007, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[3]  Shunichi Takeda,et al.  Single-stranded DNA-binding protein hSSB1 is critical for genomic stability , 2008, Nature.

[4]  S. Jackson,et al.  Regulation of DNA-damage responses and cell-cycle progression by the chromatin remodelling factor CHD4 , 2010, The EMBO journal.

[5]  S. Elledge,et al.  Ubc13/Rnf8 ubiquitin ligases control foci formation of the Rap80/Abraxas/Brca1/Brcc36 complex in response to DNA damage , 2007, Proceedings of the National Academy of Sciences.

[6]  J. Ellenberg,et al.  RNF168 Binds and Amplifies Ubiquitin Conjugates on Damaged Chromosomes to Allow Accumulation of Repair Proteins , 2009, Cell.

[7]  R. Abraham.  Cell cycle checkpoint signaling through the ATM and ATR kinases. , 2001, Genes & development.

[8]  S. Schreiber,et al.  Molecular association between ATR and two components of the nucleosome remodeling and deacetylating complex, HDAC2 and CHD4. , 1999, Biochemistry.

[9]  Y. Shiloh,et al.  Nuclear retention of ATM at sites of DNA double strand breaks. , 2001, The Journal of biological chemistry.

[10]  Yair Andegeko,et al.  Requirement of the MRN complex for ATM activation by DNA damage , 2003, The EMBO journal.

[11]  S. Schreiber,et al.  Chromatin deacetylation by an ATP-dependent nucleosome remodelling complex , 1998, Nature.

[12]  G. Stewart.  Solving the RIDDLE of 53BP1 recruitment to sites of damage , 2009, Cell cycle.

[13]  B. A. Ballif,et al.  ATM and ATR Substrate Analysis Reveals Extensive Protein Networks Responsive to DNA Damage , 2007, Science.

[14]  Edward S. Miller,et al.  The RIDDLE Syndrome Protein Mediates a Ubiquitin-Dependent Signaling Cascade at Sites of DNA Damage , 2009, Cell.

[15]  Ricky W. Johnstone,et al.  Epigenetics in cancer: Targeting chromatin modifications , 2009, Molecular Cancer Therapeutics.

[16]  T. Misteli,et al.  The emerging role of nuclear architecture in DNA repair and genome maintenance , 2009, Nature Reviews Molecular Cell Biology.

[17]  S. Elledge,et al.  Linkage of ATM to cell cycle regulation by the Chk2 protein kinase. , 1998, Science.

[18]  K. Khanna,et al.  DNA double-strand breaks: signaling, repair and the cancer connection , 2001, Nature Genetics.

[19]  David J. Chen,et al.  hSSB1 rapidly binds at the sites of DNA double-strand breaks and is required for the efficient recruitment of the MRN complex , 2010, Nucleic acids research.

[20]  N. L. La Thangue,et al.  Histone deacetylase inhibitors open new doors in cancer therapy. , 2004, Biochemical pharmacology.

[21]  Jiri Bartek,et al.  RNF8 Ubiquitylates Histones at DNA Double-Strand Breaks and Promotes Assembly of Repair Proteins , 2007, Cell.

[22]  Y Taya,et al.  Activation of the ATM kinase by ionizing radiation and phosphorylation of p53. , 1998, Science.

[23]  K. Khanna,et al.  ATM, a central controller of cellular responses to DNA damage , 2001, Cell Death and Differentiation.

[24]  R. Greenberg,et al.  ATM-Dependent Chromatin Changes Silence Transcription In cis to DNA Double-Strand Breaks , 2010, Cell.

[25]  Weidong Wang,et al.  NURD, a novel complex with both ATP-dependent chromatin-remodeling and histone deacetylase activities. , 1998, Molecular cell.

[26]  Binding of the CHD4 PHD2 finger to histone H3 is modulated by covalent modifications. , 2009, The Biochemical journal.

[27]  T. Misteli,et al.  Aging-related chromatin defects via loss of the NURD complex , 2009, Nature Cell Biology.

[28]  Michael B. Yaffe,et al.  RNF8 Transduces the DNA-Damage Signal via Histone Ubiquitylation and Checkpoint Protein Assembly , 2007, Cell.

[29]  A. Wolffe,et al.  A multiple subunit Mi-2 histone deacetylase from Xenopus laevis cofractionates with an associated Snf2 superfamily ATPase , 1998, Current Biology.

[30]  G. Längst,et al.  The dMi‐2 chromodomains are DNA binding modules important for ATP‐dependent nucleosome mobilization , 2002, The EMBO journal.

[31]  W. Bin,et al.  Ubc13/Rnf8ユビキチンリガーゼはDNS損傷に反応してRap80/Abraxas/Brca1/Brca1/Brcc36複合体のフォーカス形成を制御する , 2007 .

[32]  Y Taya,et al.  Enhanced phosphorylation of p53 by ATM in response to DNA damage. , 1998, Science.

[33]  M. Gatei,et al.  ATM-dependent phosphorylation of nibrin in response to radiation exposure , 2000, Nature Genetics.

[34]  Laurence Pelletier,et al.  Orchestration of the DNA-Damage Response by the RNF8 Ubiquitin Ligase , 2007, Science.

[35]  P. Ménard,et al.  The chromatin-remodeling factor CHD4 coordinates signaling and repair after DNA damage , 2010, The Journal of cell biology.