Tip60-mediated acetylation activates transcription independent apoptotic activity of Abl

BackgroundThe proto-oncogene, c-Abl encodes a ubiquitously expressed tyrosine kinase that critically governs the cell death response induced by genotoxic agents such as ionizing radiation and cisplatin. The catalytic function of Abl, which is essential for executing DNA damage response (DDR), is normally tightly regulated but upregulated several folds upon IR exposure due to ATM-mediated phosphorylation on S465. However, the mechanism/s leading to activation of Abl's apoptotic activity is currently unknown.ResultsWe investigated the role of acetyl modification in regulating apoptotic activity of Abl and the results showed that DNA strand break-inducing agents, ionizing radiation and bleomycin induced Abl acetylation. Using mass spectrophotometry and site-specific acetyl antibody, we identified Abl K921, located in the DNA binding domain, and conforming to one of the lysine residue in the consensus acetylation motif (K XXK--X3-5--SGS) is acetylated following DNA damage. We further observed that the S465 phosphorylated Abl is acetyl modified during DNA damage. Signifying the modification, cells expressing the non acetylatable K921R mutant displayed attenuated apoptosis compared to wild-type in response to IR or bleomycin treatment. WT-Abl induced apoptosis irrespective of new protein synthesis. Furthermore, upon γ-irradiation K921R-Abl displayed reduced chromatin binding compared to wild type. Finally, loss of Abl K921 acetylation in Tip60-knocked down cells and co-precipitation of Abl with Tip60 in DNA damaged cells identified Tip60 as an Abl acetylase.ConclusionCollective data showed that DNA damage-induced K921 Abl acetylation, mediated by Tip60, stimulates transcriptional-independent apoptotic activity and chromatin-associative property thereby defining a new regulatory mechanism governing Abl's DDR function.

[1]  R. Weichselbaum,et al.  Role for c-Abl tyrosine kinase in growth arrest response to DNA damage , 1996, Nature.

[2]  B. Amati,et al.  Tip60 in DNA damage response and growth control: many tricks in one HAT. , 2006, Trends in cell biology.

[3]  Tohru Natsume,et al.  JNK phosphorylation of 14-3-3 proteins regulates nuclear targeting of c-Abl in the apoptotic response to DNA damage , 2005, Nature Cell Biology.

[4]  David J. Chen,et al.  Suppression of nonhomologous end joining repair by overexpression of HMGA2. , 2009, Cancer research.

[5]  Guoming Sun,et al.  c-Abl Kinase Regulates Curcumin-Induced Cell Death through Activation of c-Jun N-Terminal Kinase , 2007, Molecular Pharmacology.

[6]  M. Štros.  HMGB proteins: interactions with DNA and chromatin. , 2010, Biochimica et biophysica acta.

[7]  G. Blandino,et al.  DNA damage-dependent acetylation of p73 dictates the selective activation of apoptotic target genes. , 2002, Molecular cell.

[8]  T. T. Su.  Cellular responses to DNA damage: one signal, multiple choices. , 2006, Annual review of genetics.

[9]  Y. Shaul,et al.  Role of c-Abl in the DNA damage stress response , 2005, Cell Research.

[10]  D. Kufe,et al.  c-Abl Tyrosine Kinase Regulates the Human Rad9 Checkpoint Protein in Response to DNA Damage , 2002, Molecular and Cellular Biology.

[11]  Jean Y. J. Wang,et al.  Binding of A/T-rich DNA by Three High Mobility Group-like Domains in c-Abl Tyrosine Kinase* , 1996, The Journal of Biological Chemistry.

[12]  P. Sung,et al.  Regulation of Rad51 Function by c-Abl in Response to DNA Damage* , 1998, The Journal of Biological Chemistry.

[13]  Jasmine Y. Young,et al.  DNA bending by the mammalian high-mobility group protein AT hook 2. , 2010, Biochemistry.

[14]  R. Bronson,et al.  Neonatal lethality and lymphopenia in mice with a homozygous disruption of the c-abl proto-oncogene , 1991, Cell.

[15]  T. Pandita,et al.  Regulation of DNA-dependent Protein Kinase Activity by Ionizing Radiation-activated Abl Kinase Is an ATM-dependent Process* , 2000, The Journal of Biological Chemistry.

[16]  Bruce Stillman,et al.  Chromatin Association of Human Origin Recognition Complex, Cdc6, and Minichromosome Maintenance Proteins during the Cell Cycle: Assembly of Prereplication Complexes in Late Mitosis , 2000, Molecular and Cellular Biology.

[17]  M. Nissen,et al.  The A.T-DNA-binding domain of mammalian high mobility group I chromosomal proteins. A novel peptide motif for recognizing DNA structure. , 1990, The Journal of biological chemistry.

[18]  S. Rea,et al.  Males absent on the first (MOF): from flies to humans , 2007, Oncogene.

[19]  Hidde Ploegh,et al.  Acetylation of the C terminus of Ku70 by CBP and PCAF controls Bax-mediated apoptosis. , 2004, Molecular cell.

[20]  R. Weichselbaum,et al.  Determination of cell fate by c-Abl activation in the response to DNA damage , 1998, Oncogene.

[21]  J. Wang,et al.  Identification of a binding site in c-Ab1 tyrosine kinase for the C-terminal repeated domain of RNA polymerase II , 1996, Molecular and cellular biology.

[22]  A. Gulino,et al.  Specific Role for p300/CREB-binding Protein-associated Factor Activity in E2F1 Stabilization in Response to DNA Damage* , 2004, Journal of Biological Chemistry.

[23]  S. Chin,et al.  p300 regulates p53-dependent apoptosis after DNA damage in colorectal cancer cells by modulation of PUMA/p21 levels. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[24]  T. Ludwig,et al.  Involvement of Human MOF in ATM Function , 2005, Molecular and Cellular Biology.

[25]  Junjie Chen,et al.  The DNA damage response pathways: at the crossroad of protein modifications , 2008, Cell Research.

[26]  J. Bartek,et al.  The DNA-damage response in human biology and disease , 2009, Nature.

[27]  W. Gu,et al.  Functional Analysis of the Roles of Posttranslational Modifications at the p53 C Terminus in Regulating p53 Stability and Activity , 2005, Molecular and Cellular Biology.

[28]  D. Srinivasan,et al.  Aggressive breast cancer cells are dependent on activated Abl kinases for proliferation, anchorage-independent growth and survival , 2008, Oncogene.

[29]  Ronen Marmorstein,et al.  Acetylation of the p53 DNA-binding domain regulates apoptosis induction. , 2006, Molecular cell.

[30]  D. Trouche,et al.  To die or not to die: a HAT trick. , 2006, Molecular cell.

[31]  Xuan Liu,et al.  Stimulation of p53 DNA Binding by c-Abl Requires the p53 C Terminus and Tetramerization , 2000, Molecular and Cellular Biology.

[32]  W. Gu,et al.  Acetylation of p53 augments its site-specific DNA binding both in vitro and in vivo. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Jean Y. J. Wang.  Regulation of cell death by the Abl tyrosine kinase , 2000, Oncogene.

[34]  H. Kitao,et al.  Regulation of Ionizing Radiation-induced Rad52 Nuclear Foci Formation by c-Abl-mediated Phosphorylation* , 2002, The Journal of Biological Chemistry.

[35]  D. Baltimore,et al.  Ataxia telangiectasia mutant protein activates c-Abl tyrosine kinase in response to ionizing radiation , 1997, Nature.

[36]  Kelvin K. Tsai,et al.  c-Abl stabilizes p73 by a phosphorylation-augmented interaction. , 2003, Cancer research.

[37]  Jacques Côté,et al.  The diverse functions of histone acetyltransferase complexes. , 2003, Trends in genetics : TIG.

[38]  D. Sgroi,et al.  The p63/p73 network mediates chemosensitivity to cisplatin in a biologically defined subset of primary breast cancers. , 2007, The Journal of clinical investigation.

[39]  Xiaofeng Jiang,et al.  A role for the Tip60 histone acetyltransferase in the acetylation and activation of ATM. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[40]  T. Holak,et al.  c-Abl Phosphorylates Hdmx and Regulates Its Interaction with p53* , 2009, Journal of Biological Chemistry.

[41]  B. Corfe,et al.  Sp1 acetylation is associated with loss of DNA binding at promoters associated with cell cycle arrest and cell death in a colon cell line , 2010, Molecular Cancer.

[42]  S. Goff,et al.  Mice homozygous for the abl m1 mutation show poor viability and depletion of selected B and T cell populations , 1991, Cell.

[43]  T. Uchiumi,et al.  Tip60 promotes prostate cancer cell proliferation by translocation of androgen receptor into the nucleus , 2010, The Prostate.

[44]  C. Bailly,et al.  The DNA binding domain of the human c-Abl tyrosine kinase preferentially binds to DNA sequences containing an AAC motif and to distorted DNA structures. , 1998, Biochemistry.

[45]  Jean Y. J. Wang,et al.  Cell Cycle—Regulated Binding of c-Abl Tyrosine Kinase to DNA , 1992, Science.

[46]  I. Hickson,et al.  Cellular responses to DNA damage. , 2001, Annual review of pharmacology and toxicology.

[47]  D. McDonald,et al.  Nuclear-cytoplasmic shuttling of C-ABL tyrosine kinase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[48]  S. Gonfloni.  DNA damage stress response in germ cells: role of c-Abl and clinical implications , 2010, Oncogene.