c-Abl Tyrosine Kinase Regulates Caspase-9 Autocleavage in the Apoptotic Response to DNA Damage

Activation of the initiator caspase-9 is essential for induction of apoptosis by developmental signals, oncogenic transformation, and genotoxic stress. The c-Abl tyrosine kinase is also involved in the apoptotic response to DNA damage. The present results demonstrate that c-Abl binds directly to caspase-9. We show that c-Abl phosphorylates caspase-9 on Tyr-153 in vitro and in cells treated with DNA damaging agents. Moreover, inhibition of c-Abl with STI571 blocked DNA damage-induced autoprocessing of caspase-9 to the p35 subunit and activation of caspase-3. Caspase-9(Y153F) also attenuated DNA damage-induced processing of caspase-9 to p35, activation of caspase-3, and apoptosis. These findings indicate that caspase-9 autoprocessing is regulated by c-Abl in the apoptotic response to genotoxic stress. Activation of the initiator caspase-9 is essential for induction of apoptosis by developmental signals, oncogenic transformation, and genotoxic stress. The c-Abl tyrosine kinase is also involved in the apoptotic response to DNA damage. The present results demonstrate that c-Abl binds directly to caspase-9. We show that c-Abl phosphorylates caspase-9 on Tyr-153 in vitro and in cells treated with DNA damaging agents. Moreover, inhibition of c-Abl with STI571 blocked DNA damage-induced autoprocessing of caspase-9 to the p35 subunit and activation of caspase-3. Caspase-9(Y153F) also attenuated DNA damage-induced processing of caspase-9 to p35, activation of caspase-3, and apoptosis. These findings indicate that caspase-9 autoprocessing is regulated by c-Abl in the apoptotic response to genotoxic stress. Caspase-9 is the initiator caspase of the apoptosome, an oligomeric complex that controls the intrinsic apoptotic pathway. Formation of the apoptosome is induced by release of mitochondrial cytochrome c into the cytosol. Cytochrome c associates with Apaf-1 and thereby promotes its oligomerization and recruitment of caspase-9 (1Adrain C. Slee E. Harte M. Martin S. J. Biol. Chem. 1999; 274: 20855-20860Abstract Full Text Full Text PDF PubMed Scopus (98) Google Scholar, 2Zou H. Li X. Liu X. Wang X. J. Biol. Chem. 1999; 274: 11549-11556Abstract Full Text Full Text PDF PubMed Scopus (1794) Google Scholar, 3Jiang X. Wang X. J. Biol. Chem. 2000; 275: 31199-31203Abstract Full Text Full Text PDF PubMed Scopus (414) Google Scholar). Binding to Apaf-1 increases activity of the caspase-9 protease and autocleavage of the p46 pro-caspase-9 at Asp-315 to yield p35 and p12 subunits (4Srinivasula S. Ahmad M. Alnemri T. Alnemri E. Mol. Cell. 1998; 1: 949-957Abstract Full Text Full Text PDF PubMed Scopus (963) Google Scholar, 5Stennicke H. Deveraux Q. Humke E. Reed J. Dixit V. Salvesen G. J. Biol. Chem. 1999; 26: 8359-8362Abstract Full Text Full Text PDF Scopus (418) Google Scholar, 6Rodriguez J. Lazebnik Y. Genes Dev. 1999; 13: 3179-3184Crossref PubMed Scopus (458) Google Scholar). Caspase-9 activation requires interaction with the Apaf-1 caspase recruitment domain, an increase in local concentrations of caspase-9, and the formation of caspase-9 dimers (7Renatus M. Stennicke H.R. Scott F.L. Liddington R.C. Salvesen G.S. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 14250-14255Crossref PubMed Scopus (367) Google Scholar, 8Shiozaki E.N. Chai J. Shi Y. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: 4197-4202Crossref PubMed Scopus (111) Google Scholar). Following autoprocessing in the apoptosome, caspase-9 cleaves and activates caspase-3. In turn, caspase-3 directs feedback cleavage of caspase-9 at Asp-330 to generate p37 and p10 subunits (4Srinivasula S. Ahmad M. Alnemri T. Alnemri E. Mol. Cell. 1998; 1: 949-957Abstract Full Text Full Text PDF PubMed Scopus (963) Google Scholar, 9Slee E.A. Harte M.T. Kluck R.M. Wolf B.B. Casiano C.A. Newmeyer D.D. Wang H.G. Reed J.C. Nicholson D.W. Alnemri E.S. Green D.R. Martin S.J. J. Cell Biol. 1999; 144: 281-292Crossref PubMed Scopus (1679) Google Scholar). The caspase-9 p12, and not the p10, subunit contains four N-terminal amino acids that bind to the third baculoviral repeat of the X-linked inhibitor of apoptosis (10Srinivasula S.M. Hegde R. Saleh A. Datta P. Shiozaki E. Chai J. Lee R.A. Robbins P.D. Fernandes-Alnemri T. Shi Y. Alnemri E.S. Nature. 2001; 410: 112-116Crossref PubMed Scopus (859) Google Scholar), which maintains caspase-9 in the inactive monomer conformation (11Fesik S.W. Shi Y. Science. 2001; 294: 1477-1478Crossref PubMed Scopus (139) Google Scholar, 12Shiozaki E.N. Chai J. Rigotti D.J. Riedl S.J. Li P. Srinivasula S.M. Alnemri E.S. Fairman R. Shi Y. Mol. Cell. 2003; 11: 519-527Abstract Full Text Full Text PDF PubMed Scopus (573) Google Scholar) and functions as a tether for caspase-3 (13Bratton S.B. Walker G. Srinivasula S.M. Sun X.M. Butterworth M. Alnemri E.S. Cohen G.M. EMBO J. 2001; 20: 998-1009Crossref PubMed Scopus (337) Google Scholar, 14Hill M.M. Adrain C. Duriez P.J. Creagh E.M. Martin S.J. EMBO J. 2004; 23: 2134-2145Crossref PubMed Scopus (217) Google Scholar). Other studies have demonstrated that caspase-9 activity is inhibited by Akt-mediated phosphorylation on Ser-196 (15Cardone M. Roy N. Stennicke H. Salvesen G. Franke T. Stanbridge E. Frisch S. Reed J. Science. 1998; 282: 1318-1321Crossref PubMed Scopus (2728) Google Scholar) and by extracellular signal-regulated kinase-mediated phosphorylation on Thr-125 (16Allan L.A. Morrice N. Brady S. Magee G. Pathak S. Clarke P.R. Nat. Cell Biol. 2003; 5: 647-654Crossref PubMed Scopus (395) Google Scholar). However, it is not known whether phosphorylation of caspase-9 contributes to autocleavage of this important apoptotic initiator. The c-Abl tyrosine kinase is activated in the response of cells to genotoxic stress (17Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (460) Google Scholar). The product of the gene mutated in ataxia telangiectasia is responsible in part for c-Abl activation (18Shafman T. Khanna K.K. Kedar P. Spring K. Kozlov S. Yen T. Hobson K. Gatei M. Zhang N. Watters D. Egerton M. Shiloh Y. Kharbanda S. Kufe D. Lavin M.F. Nature. 1997; 387: 520-523Crossref PubMed Scopus (419) Google Scholar, 19Baskaran R. Wood L.D. Whitaker L.L. Xu Y. Barlow C. Canman C.E. Morgan S.E. Baltimore D. Wynshaw-Boris A. Kastan M.B. Wang J.Y.J. Nature. 1997; 387: 516-519Crossref PubMed Scopus (485) Google Scholar). Other work has demonstrated that nuclear c-Abl interacts with the DNA-dependent protein kinase (DNA-PK)-Ku complex (20Kharbanda S. Pandey P. Jin S. Inoue S. Bharti A. Yuan Z.-M. Weichselbaum R. Weaver D. Kufe D. Nature. 1997; 386: 732-735Crossref PubMed Scopus (237) Google Scholar, 21Jin S. Kharbanda S. Mayer B. Kufe D. Weaver D.T. J. Biol. Chem. 1997; 272: 24763-24766Abstract Full Text Full Text PDF PubMed Scopus (91) Google Scholar). Phosphorylation of c-Abl by the catalytic subunit DNA-PKcs stimulates c-Abl activity (20Kharbanda S. Pandey P. Jin S. Inoue S. Bharti A. Yuan Z.-M. Weichselbaum R. Weaver D. Kufe D. Nature. 1997; 386: 732-735Crossref PubMed Scopus (237) Google Scholar). Activation of c-Abl by DNA damage or inhibition of DNA replication contributes to the induction of apoptosis by mechanisms in part dependent on the p53 tumor suppressor and its homolog p73 (22Yuan Z. Huang Y. Fan M. Sawers C. Kharbanda S. Kufe D. J. Biol. Chem. 1996; 271: 26457-26460Abstract Full Text Full Text PDF PubMed Scopus (64) Google Scholar, 23Yuan Z. Huang Y. Ishiko T. Kharbanda S. Weichselbaum R. Kufe D. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 1437-1440Crossref PubMed Scopus (178) Google Scholar, 24Yuan Z.M. Shioya H. Ishiko T. Sun X. Huang Y. Lu H. Kharbanda S. Weichselbaum R. Kufe D. Nature. 1999; 399: 814-817Crossref PubMed Scopus (539) Google Scholar, 25Agami R. Blandino G. Oren M. Shaul Y. Nature. 1999; 399: 809-813Crossref PubMed Scopus (504) Google Scholar, 26Gong J. Costanzo A. Yang H. Melino G. Kaelin Jr., W. Levrero M. Wang J.Y.J. Nature. 1999; 399: 806-809Crossref PubMed Scopus (833) Google Scholar). c-Abl also contributes to DNA damage-induced activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) kinase-1, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase pathways (17Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (460) Google Scholar, 27Kharbanda S. Pandey P. Ren R. Feller S. Mayer B. Zon L. Kufe D. J. Biol. Chem. 1995; 270: 30278-30281Abstract Full Text Full Text PDF PubMed Scopus (127) Google Scholar, 28Pandey P. Raingeaud J. Kaneki M. Weichselbaum R. Davis R. Kufe D. Kharbanda S. J. Biol. Chem. 1996; 271: 23775-23779Abstract Full Text Full Text PDF PubMed Scopus (119) Google Scholar, 29Kharbanda S. Pandey P. Yamauchi T. Kumar S. Kaneki M. Kumar V. Bharti A. Yuan Z. Ghanem L. Rana A. Weichselbaum R. Johnson G. Kufe D. Mol. Cell. Biol. 2000; 20: 4979-4989Crossref PubMed Scopus (84) Google Scholar). Moreover, c-Abl interactions with Rad51, Rad9, and the hTERT telomerase catalytic subunit have been implicated in the apoptotic response to DNA damage (30Yuan Z.M. Huang Y. Ishiko T. Nakada S. Utsugisawa T. Kharbanda S. Sung P. Shinohara A. Weichselbaum R. Kufe D. J. Biol. Chem. 1998; 273: 3799-3802Abstract Full Text Full Text PDF PubMed Scopus (185) Google Scholar, 31Kharbanda S. Kumar V. Dhar S. Pandey P. Chen C. Majumder P. Yuan Z. Whang Y. Strauss W. Pandita T. Weaver D. Kufe D. Curr. Biol. 2000; 10: 568-575Abstract Full Text Full Text PDF PubMed Scopus (131) Google Scholar, 32Yoshida K. Komatsu K. Wang H.-G. Kufe D. Mol. Cell. Biol. 2002; 22: 3292-3300Crossref PubMed Scopus (88) Google Scholar). In concert with these studies, DNA damage-induced apoptosis is attenuated in cells that i) express a kinase-inactive, dominant-negative c-Abl(K-R) mutant, ii) are null for c-Abl (c-abl–/–), or iii) are treated with the c-Abl kinase inhibitor STI571 (23Yuan Z. Huang Y. Ishiko T. Kharbanda S. Weichselbaum R. Kufe D. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 1437-1440Crossref PubMed Scopus (178) Google Scholar, 33Huang Y. Yuan Z.M. Ishiko T. Nakada S. Utsugisawa T. Kato T. Kharbanda S. Kufe D.W. Oncogene. 1997; 15: 1947-1952Crossref PubMed Scopus (40) Google Scholar, 34Raina D. Mishra N. Kumar S. Kharbanda S. Saxena S. Kufe D. Mol. Pharmacol. 2002; 61: 1489-1495Crossref PubMed Scopus (20) Google Scholar). Notably, however, there are no known interactions between c-Abl and the initiator or effector caspases. The present studies demonstrate that c-Abl phosphorylates caspase-9 on Tyr-153 in vitro and in the response to DNA damage. We also show that c-Abl-mediated phosphorylation of caspase-9 contributes to DNA damage-induced autoprocessing of caspase-9, activation of caspase-3, and apoptosis. Cell Culture—Human U-937 myeloid leukemia cells (ATCC, Manassas, VA) were cultured in RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum, 100 units/ml penicillin, 100 μg/ml streptomycin, and 2 mm l-glutamine. Wild-type, c-abl–/– and c-abl+/– mouse fibroblasts (35Tybulewicz V.L.J. Crawford C.E. Jackson P.K. Bronson R.T. Mulligan R.C. Cell. 1991; 65: 1153-1163Abstract Full Text PDF PubMed Scopus (1159) Google Scholar) were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and antibiotics. Cells were treated with 10 μm 1-(β-d-arabinofuranosyl)cytosine (araC) 1The abbreviations used are: araC, 1-(β-d-arabinofuranosyl)cytosine; GST, glutathione S-transferase; SH3, Src homology 3; IR, ionizing radiation. (Sigma). Irradiation was performed with a γ-ray source (137Cs, Gammacell 1000; Atomic Energy of Canada, Ltd., Ontario, Canada) at a fixed dose of 13 grays/min. Plasmid Construction—The vector expressing pCDNA3-caspase-9 has been described (36Pandey P. Saleh A. Nakazawa A. Kumar S. Srinivasula S.M. Kumar V. Weichselbaum R. Nalin C. Alnemri E.S. Kufe D. Kharbanda S. EMBO J. 2000; 19: 4310-4322Crossref PubMed Scopus (486) Google Scholar). The caspase-9(Y153F) mutant was generated by site-directed mutagenesis and confirmed by DNA sequencing. Caspase-9 and caspase-9(Y153F) were subcloned into pGEX4T-1 (Amersham Biosciences) at the BamH1 and EcoR1 sites and into pLXIN (Clontech) at the BamH1 site. Retroviral Transduction—PT67 cells were transiently transfected with pLXIN or pLXIN-caspase-9(Y153F) in the presence of Lipofectamine. At 48 h after transfection, cells were selected in medium containing G418. Retroviral titers of the supernatants were determined using NIH3T3 cells. Filtered retroviral supernatants containing polybrene (Sigma) were used to infect U-937 cells. At 24 h after infection, cells were seeded into methylcellulose medium (Stem Cell Technologies) and single cell clones were selected in G418. Immunoprecipitation and Immunoblot Analysis—Cell lysates were prepared as described (37Yoshida K. Kharbanda S. Kufe D. J. Biol. Chem. 1999; 274: 34663-34668Abstract Full Text Full Text PDF PubMed Scopus (49) Google Scholar) and cleared by centrifugation at 12,000 × g for 15 min. Cytosolic S15 lysates were prepared as described (14Hill M.M. Adrain C. Duriez P.J. Creagh E.M. Martin S.J. EMBO J. 2004; 23: 2134-2145Crossref PubMed Scopus (217) Google Scholar). Soluble proteins (500 μg) were incubated with anti-caspase-9 (sc-7885; Santa Cruz Biotechnology) for 2 h at 4 °C, followed by precipitation with protein A-Sepharose beads for 1 h. Immune complexes and cell lysates (50 μg) were subjected to immunoblot analysis with anti-c-Abl (Ab-3; Oncogene Research Products), anti-caspase-9 (sc-8355; Santa Cruz) (monoclonal 96–2-22; Upstate Biotechnology Inc.), anti-phospho-Tyr (4G10; Upstate Biotechnology), anti-cytochrome c (36Pandey P. Saleh A. Nakazawa A. Kumar S. Srinivasula S.M. Kumar V. Weichselbaum R. Nalin C. Alnemri E.S. Kufe D. Kharbanda S. EMBO J. 2000; 19: 4310-4322Crossref PubMed Scopus (486) Google Scholar), anti-caspase-3 (sc-7148; Santa Cruz) and anti-β-actin (Sigma). The antigen-antibody complexes were visualized by chemiluminescence (PerkinElmer Life Sciences). In Vitro Binding Assays—Cell lysates were incubated with purified GST or GST-caspase-9 in lysis for 2 h at 4 In GST, SH3, or were incubated with purified to glutathione beads were by In Vitro and purified cells Z.M. Shioya H. Ishiko T. Sun X. Huang Y. Lu H. Kharbanda S. Weichselbaum R. Kufe D. Nature. 1999; 399: 814-817Crossref PubMed Scopus (539) Google Scholar) were incubated in kinase (50 mm 10 mm 10 mm 2 mm mm with and Life for 15 at were by and was by cells with and by of cells with DNA were determined by the DNA was performed as described R. H. K. Kufe D. J. Biol. Chem. 1997; 272: Full Text Full Text PDF PubMed Scopus Google Scholar). c-Abl with whether c-Abl associates with caspase-9, lysates U-937 cells were with Immunoblot analysis of the with anti-c-Abl demonstrated complexes of c-Abl and caspase-9 were were performed on U-937 cells treated with araC, an inhibitor of DNA replication E.M. G. M. Kufe D.W. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar) and of c-Abl activation Y. Yuan Z.M. Ishiko T. Nakada S. Utsugisawa T. Kato T. Kharbanda S. Kufe D.W. Oncogene. 1997; 15: 1947-1952Crossref PubMed Scopus (40) Google Scholar, 34Raina D. Mishra N. Kumar S. Kharbanda S. Saxena S. Kufe D. Mol. Pharmacol. 2002; 61: 1489-1495Crossref PubMed Scopus (20) Google Scholar) The of c-Abl and caspase-9 was at of and was at the U-937 cell lysates were incubated with GST or a GST-caspase-9 Immunoblot analysis of the of c-Abl to GST-caspase-9 and not GST whether the interaction is purified caspase-9 was incubated with GST proteins containing or of the with anti-caspase-9 demonstrated to c-Abl and c-Abl(K-R) the c-Abl responsible for to caspase-9, incubated with GST or a GST protein that contains the c-Abl The results show that c-Abl binds to These findings indicate that c-Abl associates with caspase-9 in the response to genotoxic stress and that the interaction is by the c-Abl c-Abl Caspase-9 on whether caspase-9 functions as a c-Abl incubated with purified or and of the by and that caspase-9 is by c-Abl and not c-Abl(K-R) the phosphorylation in were mutated to and the were as for The results demonstrated in to there was no phosphorylation of with the whether c-Abl phosphorylates caspase-9 in studies were performed with and c-abl–/– mouse and tyrosine phosphorylation of caspase-9 was in not cells of c-Abl in the c-abl–/– cells was also with and tyrosine phosphorylation of caspase-9 whether caspase-9 is on Tyr-153 in generated U-937 cells expressing an vector or caspase-9(Y153F) U-937 cells transfected with caspase-9 were not for caspase-9, was with of c-Abl and caspase-9(Y153F) In immunoblot analysis of anti-caspase-9 with anti-phospho-Tyr demonstrated tyrosine phosphorylation of caspase-9 in the cells there was tyrosine phosphorylation of caspase-9 in the cells These findings indicate that c-Abl phosphorylates caspase-9 on Tyr-153 in vitro and in the response of cells to DNA damage. Caspase-9(Y153F) or STI571 DNA of of U-937 cells with is with release of mitochondrial cytochrome c to the and activation of caspase-3 at 4 and h D. Mishra N. Kumar S. Kharbanda S. Saxena S. Kufe D. Mol. Pharmacol. 2002; 61: 1489-1495Crossref PubMed Scopus (20) Google Scholar). of cells was also with processing of caspase-9 to the p37 and p35 subunits however, processing of caspase-9 to the p35 was attenuated in the cells Moreover, activation of caspase-3 was attenuated in cells with that in cells cleavage of caspase-9 to the p37 subunit (4Srinivasula S. Ahmad M. Alnemri T. Alnemri E. Mol. Cell. 1998; 1: 949-957Abstract Full Text Full Text PDF PubMed Scopus (963) Google Scholar, 9Slee E.A. Harte M.T. Kluck R.M. Wolf B.B. Casiano C.A. Newmeyer D.D. Wang H.G. Reed J.C. Nicholson D.W. Alnemri E.S. Green D.R. Martin S.J. J. Cell Biol. 1999; 144: 281-292Crossref PubMed Scopus (1679) Google Scholar). cleavage of caspase-9(Y153F) to p37 in the of caspase-3 activation that of caspase-3 activity or caspase-9 cleavage at Asp-330 in the presence of the whether inhibition of c-Abl activity has on caspase-9 and caspase-3, U-937 cells were treated with the c-Abl to STI571 no on release of cytochrome c However, STI571 was with of cleavage of caspase-9 to the p35 subunit STI571 also attenuated activation of caspase-3 not with these analysis of anti-caspase-9 of U-937 cells demonstrated that increases the formation of complexes and that STI571 this response Moreover, STI571 attenuated the cleavage of caspase-9 to p35 These findings indicate of inhibition of the c-Abl kinase with STI571 autoprocessing of caspase-9 to the p35 Caspase-9(Y153F) DNA of the caspase-9(Y153F) mutant on apoptosis were by the of cells with of cells was with a increase in DNA and this response was attenuated in cells results were at of and in U-937 cells also to of ionizing with the induction of apoptosis S. Saxena S. K. Pandey P. Kaneki M. Wang Q. K. Chen Y. A. S. Yuan Z. J. Weichselbaum R. Nalin C. Kufe D. J. Biol. Chem. 2000; 275: Full Text Full Text PDF PubMed Scopus Google Scholar). with araC, apoptosis of cells was with that with cells of these results in that cells an attenuated apoptotic response to DNA damaging agents. an for apoptotic DNA was attenuated in U-937 cells expressing caspase-9(Y153F) results were with and cells These findings indicate that caspase-9(Y153F) DNA damage-induced apoptosis. c-Abl Caspase-9 on studies have demonstrated that and extracellular signal-regulated kinase and thereby caspase-9 processing (15Cardone M. Roy N. Stennicke H. Salvesen G. Franke T. Stanbridge E. Frisch S. Reed J. Science. 1998; 282: 1318-1321Crossref PubMed Scopus (2728) Google Scholar, L.A. Morrice N. Brady S. Magee G. Pathak S. Clarke P.R. Nat. Cell Biol. 2003; 5: 647-654Crossref PubMed Scopus (395) Google Scholar). The present work that c-Abl associates with caspase-9 in the response of cells to genotoxic stress. of of c-Abl and caspase-9 were also cells were by not In vitro studies demonstrated that the c-Abl binds directly to caspase-9. The that the c-Abl interacts with a in the caspase-9 caspase recruitment amino acids not The results also demonstrate that c-Abl phosphorylates caspase-9 on The Tyr-153 in the caspase-9 subunit the caspase recruitment on studies, Tyr-153 on the inactive caspase-9 conformation and in the conformation the (7Renatus M. Stennicke H.R. Scott F.L. Liddington R.C. Salvesen G.S. Proc. Natl. Acad. Sci. U. S. A. 2001; 98: 14250-14255Crossref PubMed Scopus (367) Google Scholar). with these caspase-9 and not caspase-9(Y153F) was to tyrosine phosphorylation in the response to DNA damage. DNA damage-induced phosphorylation of caspase-9 by a was confirmed by the of this response in cells null for c-Abl Moreover, of c-Abl in c-abl+/– cells was with tyrosine phosphorylation of caspase-9. These findings indicate that caspase-9 is by c-Abl on Tyr-153 in vitro and in the response to DNA damage. c-Abl Caspase-9 in the to DNA is activated in the response of cells to genotoxic stress (17Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (460) Google Scholar). Other studies on and c-abl+/– cells have that c-Abl is essential for DNA damage-induced apoptosis (23Yuan Z. Huang Y. Ishiko T. Kharbanda S. Weichselbaum R. Kufe D. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 1437-1440Crossref PubMed Scopus (178) Google Scholar, 33Huang Y. Yuan Z.M. Ishiko T. Nakada S. Utsugisawa T. Kato T. Kharbanda S. Kufe D.W. Oncogene. 1997; 15: 1947-1952Crossref PubMed Scopus (40) Google Scholar). Moreover, inhibition of c-Abl with STI571 has been to apoptosis D. Mishra N. Kumar S. Kharbanda S. Saxena S. Kufe D. Mol. Pharmacol. 2002; 61: 1489-1495Crossref PubMed Scopus (20) Google Scholar). c-Abl interacts with that are with the apoptotic response (17Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (460) Google Scholar, Z. Huang Y. Fan M. Sawers C. Kharbanda S. Kufe D. J. Biol. Chem. 1996; 271: 26457-26460Abstract Full Text Full Text PDF PubMed Scopus (64) Google Scholar, 23Yuan Z. Huang Y. Ishiko T. Kharbanda S. Weichselbaum R. Kufe D. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 1437-1440Crossref PubMed Scopus (178) Google Scholar, 24Yuan Z.M. Shioya H. Ishiko T. Sun X. Huang Y. Lu H. Kharbanda S. Weichselbaum R. Kufe D. Nature. 1999; 399: 814-817Crossref PubMed Scopus (539) Google Scholar, 25Agami R. Blandino G. Oren M. Shaul Y. Nature. 1999; 399: 809-813Crossref PubMed Scopus (504) Google Scholar, 26Gong J. Costanzo A. Yang H. Melino G. Kaelin Jr., W. Levrero M. Wang J.Y.J. Nature. 1999; 399: 806-809Crossref PubMed Scopus (833) Google Scholar, 27Kharbanda S. Pandey P. Ren R. Feller S. Mayer B. Zon L. Kufe D. J. Biol. Chem. 1995; 270: 30278-30281Abstract Full Text Full Text PDF PubMed Scopus (127) Google Scholar, 28Pandey P. Raingeaud J. Kaneki M. Weichselbaum R. Davis R. Kufe D. Kharbanda S. J. Biol. Chem. 1996; 271: 23775-23779Abstract Full Text Full Text PDF PubMed Scopus (119) Google Scholar, 29Kharbanda S. Pandey P. Yamauchi T. Kumar S. Kaneki M. Kumar V. Bharti A. Yuan Z. Ghanem L. Rana A. Weichselbaum R. Johnson G. Kufe D. Mol. Cell. Biol. 2000; 20: 4979-4989Crossref PubMed Scopus (84) Google Scholar, Z.M. Huang Y. Ishiko T. Nakada S. Utsugisawa T. Kharbanda S. Sung P. Shinohara A. Weichselbaum R. Kufe D. J. Biol. Chem. 1998; 273: 3799-3802Abstract Full Text Full Text PDF PubMed Scopus (185) Google Scholar, 31Kharbanda S. Kumar V. Dhar S. Pandey P. Chen C. Majumder P. Yuan Z. Whang Y. Strauss W. Pandita T. Weaver D. Kufe D. Curr. Biol. 2000; 10: 568-575Abstract Full Text Full Text PDF PubMed Scopus (131) Google Scholar, 32Yoshida K. Komatsu K. Wang H.-G. Kufe D. Mol. Cell. Biol. 2002; 22: 3292-3300Crossref PubMed Scopus (88) Google however, there has been no known interaction between c-Abl and the initiator or effector caspases. In the present studies, the of the interaction in cells is by the that inhibition of c-Abl with STI571 autoprocessing of caspase-9 to the p35 Caspase-9(Y153F) also blocked processing to the p35 subunit and caspase-3 results demonstrate that caspase-9(Y153F) apoptosis. into DNA and functions as an inhibitor of DNA replication E.M. G. M. Kufe D.W. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). The that caspase-9(Y153F) apoptosis that the c-Abl caspase-9 is of in the response to genotoxic agents. caspase-9 Tyr-153 is in sites that are to in caspase-3 and The present findings a in which caspase-9 autocleavage is regulated by a in the apoptotic response to genotoxic stress. We for with

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