The ARG Tyrosine Kinase Interacts with Siva-1 in the Apoptotic Response to Oxidative Stress

Abstract

The Abl family of mammalian nonreceptor tyrosine kinases consists of c-Abl and ARG (Abl-related gene). Certain insights are available regarding the involvement c-Abl in the response of cells to stress. ARG, however, has no known function in cell signaling. The present studies demonstrate that ARG associates with the proapoptotic Siva-1 protein. The functional significance of the ARG-Siva-1 interaction is supported by the finding that ARG is activated by oxidative stress and that this response involves ARG-mediated phosphorylation of Siva-1 on Tyr48. The proapoptotic effects of Siva-1 are accentuated in cells stably expressing ARG and are inhibited in ARG-deficient cells. Moreover, the proapoptotic effects of Siva-1 are abrogated by mutation of the Tyr48site. We also show that the apoptotic response to oxidative stress is attenuated in ARG-deficient cells and that this defect is corrected by reconstituting ARG expression. These findings support a model in which the activation of ARG by oxidative stress induces apoptosis by a Siva-1-dependent mechanism. The Abl family of mammalian nonreceptor tyrosine kinases consists of c-Abl and ARG (Abl-related gene). Certain insights are available regarding the involvement c-Abl in the response of cells to stress. ARG, however, has no known function in cell signaling. The present studies demonstrate that ARG associates with the proapoptotic Siva-1 protein. The functional significance of the ARG-Siva-1 interaction is supported by the finding that ARG is activated by oxidative stress and that this response involves ARG-mediated phosphorylation of Siva-1 on Tyr48. The proapoptotic effects of Siva-1 are accentuated in cells stably expressing ARG and are inhibited in ARG-deficient cells. Moreover, the proapoptotic effects of Siva-1 are abrogated by mutation of the Tyr48site. We also show that the apoptotic response to oxidative stress is attenuated in ARG-deficient cells and that this defect is corrected by reconstituting ARG expression. These findings support a model in which the activation of ARG by oxidative stress induces apoptosis by a Siva-1-dependent mechanism. The mammalian c-Abl and ARG 1The abbreviations used are:ARGAbl-related geneDNA-PKDNA-dependent protein kinaseROSreactive oxygen speciesPKCprotein kinase CGSTglutathioneS-transferaseGFPgreen fluorescence proteinPAGEpolyacrylamide gel electrophoresisMEFmouse embryo fibroblastSH2/SH3Src homology 2 and 3 (domains) nonreceptor tyrosine kinases are ubiquitously expressed in adult tissues (1Goff S.P. Gilboa E. Witte O.N. Baltimore D. Cell. 1980; 22: 777-785Abstract Full Text PDF PubMed Scopus (279) Google Scholar, 2Kruh G.D. Perego R. Miki T. Aaronson S.A. Proc. Natl. Acad. Sci. U. S. A. 1990; 87: 5802-5806Crossref PubMed Scopus (135) Google Scholar). These proteins contain N-terminal SH3, SH2, and kinase domains that share ∼90% identity. The C-terminal regions of c-Abl and ARG share 29% identity and are distinguished from other nonreceptor tyrosine kinases by the presence of globular (G) and filamentous (F) actin-binding domains (3Van Etten R.A. Jackson P.K. Baltimore D. Sanders M.C. Matsuddaira P.T. Janmey P.A. J. Cell Biol. 1994; 124: 325-340Crossref PubMed Scopus (237) Google Scholar). The c-Abl protein is expressed in the nucleus and cytoplasm, whereas ARG has been detected predominately in the cytoplasm (4Wang B. Kruh G.D. Oncogene. 1996; 13: 193-197PubMed Google Scholar). In addition, the C-terminal region of c-Abl differs from ARG by the presence of a nuclear localization signal (5Van Etten R.A. Jackson P. Baltimore D. Cell. 1989; 58: 669-678Abstract Full Text PDF PubMed Scopus (335) Google Scholar), sites for phosphorylation by the Cdc2 kinase (6Kipreos E.T. Wang J.Y. Science. 1990; 248: 217-220Crossref PubMed Scopus (118) Google Scholar), and DNA binding sequences (7Kipreos E.T. Wang J.Y. Science. 1992; 256: 382-385Crossref PubMed Scopus (177) Google Scholar). The structural differences of the C-terminal regions have suggested that c-Abl and ARG may share only certain cellular functions.Mice with targeted disruption of the c-abl gene are born runted with head and eye abnormalities and succumb as neonates to defective lymphopoiesis (8Tybulewicz 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 (1158) Google Scholar, 9Schwartzberg P.L. Stall A.M. Hardin J.D. Bowdish K.S. Humaran T. Boast S. Harbison M.L. Robertson E.J. Goff S.P. Cell. 1991; 65: 1165-1175Abstract Full Text PDF PubMed Scopus (299) Google Scholar). Mice deficient in ARG develop normally but exhibit behavioral abnormalities (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar). Embryos deficient in both c-Abl and ARG exhibit defects in neurolation and die before 11 days postcoitum (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar). These findings and the observation thatabl−/−,arg−/− neuroepithelial cells exhibit an altered actin cytoskeleton have supported roles for c-Abl and ARG in the regulation of actin microfilaments (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar).Other studies have demonstrated that c-Abl is involved in the cellular response to stress (11Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (458) Google Scholar). Nuclear c-Abl associates with the DNA-dependent protein kinase (DNA-PK) complex (12Kharbanda 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, 13Jin 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) and with the product of the gene mutated in ataxia telangiectasia (14Shafman 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, 15Baskaran 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). Activation of c-Abl by DNA-PK and ataxia telangiectasia mutated gene product in cells exposed to to DNA apoptosis by in on and (11Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (458) Google Scholar, Y. Y. C. Weichselbaum R. Kharbanda S. Kufe D. Nature. 1996; PubMed Scopus Google Scholar, T. Y. Kharbanda S. Weichselbaum R. Kufe D. Nature. PubMed Scopus Google Scholar, J. A. M. Wang J.Y.J. Nature. PubMed Scopus Google Scholar, R. M. Y. Nature. PubMed Scopus Google Scholar). In the cellular response to oxygen the of c-Abl is activated by protein kinase R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). Activation of c-Abl by that of and apoptosis P. S. Weichselbaum R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google functional has been to ARG as a cell The present studies demonstrate that ARG with the proapoptotic Siva-1 protein Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar, Y. Y. Oncogene. PubMed Scopus Google Scholar). ARG Siva-1 in the cellular response to oxidative stress and induces apoptosis by a Siva-1-dependent the findings in the that ARG associates with the Siva-1 protein from cells expressing ARG and Siva-1 to with of the by with demonstrated the presence of ARG-Siva-1 of from cells expressing by with support for binding of ARG and Siva-1 from cells expressing with a protein. of the with the binding of ARG and Siva-1 studies with proteins from the ARG and ARG domains demonstrated that both binding to Siva-1 binding of SH2, but SH3, to the protein but a for binding to ARG however, the is in Siva-1 Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Y. Y. Oncogene. PubMed Scopus Google Scholar). In this SH2, and SH3, to Siva-1 These findings demonstrate that the ARG with Siva-1 and of both and and that binding of the ARG is also with Siva-1 is a for ARG, with ARG in the presence of of the by and demonstrated phosphorylation of Siva-1 a no phosphorylation Siva-1 with in which in the binding mutated to are tyrosine phosphorylation sites in Siva-1 that are and of sites to and of the proteins with ARG demonstrated of phosphorylation with but with ARG Siva-1 in with ARG and by with The show that Siva-1 is by ARG in cells no tyrosine phosphorylation of this with also by ARG, no tyrosine phosphorylation of In with findings and the presence of in of with ARG, but also in tyrosine phosphorylation These findings support for ARG-mediated phosphorylation of Siva-1 and on and in ARG, c-Abl R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, P. S. Weichselbaum R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), is activated by cells expressing with of for phosphorylation of demonstrated of ARG a the to In studies of and of in phosphorylation of the cells with however, with tyrosine phosphorylation of no phosphorylation of in These findings demonstrate that activation of ARG by is with phosphorylation of Siva-1 on of Siva-1 in response to oxidative and with used as a by and cells with ARG and to with and cells to the cells with the of for 2 to ARG kinase Siva-1 as expressing with for 2 by with and the functional significance of the ARG-Siva-1 cells that stably ARG the no of ARG on cell cell of Siva-1 in the cells with the of apoptotic cells DNA apoptosis in of Siva-1 on cells a of in apoptosis in cells as with that with and no on the and cells Siva-1 expressed in the The demonstrate Siva-1 induces apoptosis in of Siva-1 in the of ARG In with the finding that ARG Siva-1 on the of on the of whereas the proapoptotic effects of to with Siva-1 These findings demonstrate that apoptosis is on the ARG kinase function and that ARG-mediated phosphorylation of Siva-1 on is a proapoptotic Siva-1 in the apoptotic response to oxidative and cells to cells for DNA with DNA are in and cells are in the and cells in the to with the with Siva-1 for The cells by and and with cells for DNA The are expressed as the S.E. of in of cells with the findings demonstrate that ARG is activated by apoptosis in the and cells. The demonstrate that the apoptotic effects of are attenuated in as with cells however, a in apoptosis in cells The apoptotic effects of attenuated as with Moreover, of ARG in cells corrected the defect in apoptosis response to oxidative stress the cells with for 2 and for In cells by with a expressing ARG as by for DNA The are expressed as the S.E. of in of cells with ARG tyrosine kinase been with the of S. A. M. P. A. PubMed Google Scholar, Y. T. T. M. M. N. K. S. Y. Y. Google Scholar). the to c-Abl and a in neurolation as (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar), a function for ARG in cell has studies have that Abl family kinases cellular with the cytoskeleton Cell. 1989; 58: Full Text PDF PubMed Scopus Google K.K. PubMed Scopus Google Scholar). The of actin-binding domains in the C-terminal region of ARG (3Van Etten R.A. Jackson P.K. Baltimore D. Sanders M.C. Matsuddaira P.T. Janmey P.A. J. Cell Biol. 1994; 124: 325-340Crossref PubMed Scopus (237) Google Scholar) and localization of ARG with actin microfilaments (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar) have supported a in regulation of the actin The present for involvement of ARG in the cellular response to oxidative stress. Moreover, the of apoptosis by oxidative stress is attenuated in ARG-deficient cells. These findings in to the actin ARG in that an apoptotic with of the family and induces apoptosis in cells Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar). Siva-1 has also been in the of apoptosis A. K. A. R. T. J. PubMed Scopus Google Scholar). Siva-1 but which sequences by induces the apoptotic response Y. Y. Oncogene. PubMed Scopus Google Scholar). The present studies demonstrate that ARG both Siva-1 and the interaction ARG and but is with the of also demonstrate that mutation of the Siva-1 the apoptotic function of Siva-1 and that apoptosis by Siva-1 is on of These findings ARG as an of Siva-1 in the apoptotic response to oxidative stress. The mammalian c-Abl and ARG 1The abbreviations used are:ARGAbl-related geneDNA-PKDNA-dependent protein kinaseROSreactive oxygen speciesPKCprotein kinase CGSTglutathioneS-transferaseGFPgreen fluorescence proteinPAGEpolyacrylamide gel electrophoresisMEFmouse embryo fibroblastSH2/SH3Src homology 2 and 3 (domains) nonreceptor tyrosine kinases are ubiquitously expressed in adult tissues (1Goff S.P. Gilboa E. Witte O.N. Baltimore D. Cell. 1980; 22: 777-785Abstract Full Text PDF PubMed Scopus (279) Google Scholar, 2Kruh G.D. Perego R. Miki T. Aaronson S.A. Proc. Natl. Acad. Sci. U. S. A. 1990; 87: 5802-5806Crossref PubMed Scopus (135) Google Scholar). These proteins contain N-terminal SH3, SH2, and kinase domains that share ∼90% identity. The C-terminal regions of c-Abl and ARG share 29% identity and are distinguished from other nonreceptor tyrosine kinases by the presence of globular (G) and filamentous (F) actin-binding domains (3Van Etten R.A. Jackson P.K. Baltimore D. Sanders M.C. Matsuddaira P.T. Janmey P.A. J. Cell Biol. 1994; 124: 325-340Crossref PubMed Scopus (237) Google Scholar). The c-Abl protein is expressed in the nucleus and cytoplasm, whereas ARG has been detected predominately in the cytoplasm (4Wang B. Kruh G.D. Oncogene. 1996; 13: 193-197PubMed Google Scholar). In addition, the C-terminal region of c-Abl differs from ARG by the presence of a nuclear localization signal (5Van Etten R.A. Jackson P. Baltimore D. Cell. 1989; 58: 669-678Abstract Full Text PDF PubMed Scopus (335) Google Scholar), sites for phosphorylation by the Cdc2 kinase (6Kipreos E.T. Wang J.Y. Science. 1990; 248: 217-220Crossref PubMed Scopus (118) Google Scholar), and DNA binding sequences (7Kipreos E.T. Wang J.Y. Science. 1992; 256: 382-385Crossref PubMed Scopus (177) Google Scholar). The structural differences of the C-terminal regions have suggested that c-Abl and ARG may share only certain cellular gene DNA-dependent protein kinase oxygen protein kinase fluorescence protein gel embryo homology 2 and 3 (domains) Mice with targeted disruption of the c-abl gene are born runted with head and eye abnormalities and succumb as neonates to defective lymphopoiesis (8Tybulewicz 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 (1158) Google Scholar, 9Schwartzberg P.L. Stall A.M. Hardin J.D. Bowdish K.S. Humaran T. Boast S. Harbison M.L. Robertson E.J. Goff S.P. Cell. 1991; 65: 1165-1175Abstract Full Text PDF PubMed Scopus (299) Google Scholar). Mice deficient in ARG develop normally but exhibit behavioral abnormalities (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar). Embryos deficient in both c-Abl and ARG exhibit defects in neurolation and die before 11 days postcoitum (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar). These findings and the observation thatabl−/−,arg−/− neuroepithelial cells exhibit an altered actin cytoskeleton have supported roles for c-Abl and ARG in the regulation of actin microfilaments (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar). studies have demonstrated that c-Abl is involved in the cellular response to stress (11Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (458) Google Scholar). Nuclear c-Abl associates with the DNA-dependent protein kinase (DNA-PK) complex (12Kharbanda 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, 13Jin 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) and with the product of the gene mutated in ataxia telangiectasia (14Shafman 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, 15Baskaran 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). Activation of c-Abl by DNA-PK and ataxia telangiectasia mutated gene product in cells exposed to to DNA apoptosis by in on and (11Kharbanda S. Ren R. Pandey P. Shafman T.D. Feller S.M. Weichselbaum R.R. Kufe D.W. Nature. 1995; 376: 785-788Crossref PubMed Scopus (458) Google Scholar, Y. Y. C. Weichselbaum R. Kharbanda S. Kufe D. Nature. 1996; PubMed Scopus Google Scholar, T. Y. Kharbanda S. Weichselbaum R. Kufe D. Nature. PubMed Scopus Google Scholar, J. A. M. Wang J.Y.J. Nature. PubMed Scopus Google Scholar, R. M. Y. Nature. PubMed Scopus Google Scholar). In the cellular response to oxygen the of c-Abl is activated by protein kinase R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). Activation of c-Abl by that of and apoptosis P. S. Weichselbaum R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). functional has been to ARG as a cell The present studies demonstrate that ARG with the proapoptotic Siva-1 protein Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar, Y. Y. Oncogene. PubMed Scopus Google Scholar). ARG Siva-1 in the cellular response to oxidative stress and induces apoptosis by a Siva-1-dependent mechanism. the findings in the that ARG associates with the Siva-1 protein from cells expressing ARG and Siva-1 to with of the by with demonstrated the presence of ARG-Siva-1 of from cells expressing by with support for binding of ARG and Siva-1 from cells expressing with a protein. of the with the binding of ARG and Siva-1 studies with proteins from the ARG and ARG domains demonstrated that both binding to Siva-1 binding of SH2, but SH3, to the protein but a for binding to ARG however, the is in Siva-1 Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Y. Y. Oncogene. PubMed Scopus Google Scholar). In this SH2, and SH3, to Siva-1 These findings demonstrate that the ARG with Siva-1 and of both and and that binding of the ARG is also with Siva-1 is a for ARG, with ARG in the presence of of the by and demonstrated phosphorylation of Siva-1 a no phosphorylation Siva-1 with in which in the binding mutated to are tyrosine phosphorylation sites in Siva-1 that are and of sites to and of the proteins with ARG demonstrated of phosphorylation with but with ARG Siva-1 in with ARG and by with The show that Siva-1 is by ARG in cells no tyrosine phosphorylation of this with also by ARG, no tyrosine phosphorylation of In with findings and the presence of in of with ARG, but also in tyrosine phosphorylation These findings support for ARG-mediated phosphorylation of Siva-1 and on and in ARG, c-Abl R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, P. S. Weichselbaum R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), is activated by cells expressing with of for phosphorylation of demonstrated of ARG a the to In studies of and of in phosphorylation of the cells with however, with tyrosine phosphorylation of no phosphorylation of in These findings demonstrate that activation of ARG by is with phosphorylation of Siva-1 on the functional significance of the ARG-Siva-1 cells that stably ARG the no of ARG on cell cell of Siva-1 in the cells with the of apoptotic cells DNA apoptosis in of Siva-1 on cells a of in apoptosis in cells as with that with and no on the and cells Siva-1 expressed in the The demonstrate Siva-1 induces apoptosis in of Siva-1 in the of ARG In with the finding that ARG Siva-1 on the of on the of whereas the proapoptotic effects of to with Siva-1 These findings demonstrate that apoptosis is on the ARG kinase function and that ARG-mediated phosphorylation of Siva-1 on is a proapoptotic Siva-1 in the apoptotic response to oxidative and cells to cells for DNA with DNA are in and cells are in the and cells in the to with the with Siva-1 for The cells by and and with cells for DNA The are expressed as the S.E. of in of cells with the findings demonstrate that ARG is activated by apoptosis in the and cells. The demonstrate that the apoptotic effects of are attenuated in as with cells however, a in apoptosis in cells The apoptotic effects of attenuated as with Moreover, of ARG in cells corrected the defect in apoptosis response to oxidative stress the cells with for 2 and for In cells by with a expressing ARG as by for DNA The are expressed as the S.E. of in of cells with ARG tyrosine kinase been with the of S. A. M. P. A. PubMed Google Scholar, Y. T. T. M. M. N. K. S. Y. Y. Google Scholar). the to c-Abl and a in neurolation as (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar), a function for ARG in cell has studies have that Abl family kinases cellular with the cytoskeleton Cell. 1989; 58: Full Text PDF PubMed Scopus Google K.K. PubMed Scopus Google Scholar). The of actin-binding domains in the C-terminal region of ARG (3Van Etten R.A. Jackson P.K. Baltimore D. Sanders M.C. Matsuddaira P.T. Janmey P.A. J. Cell Biol. 1994; 124: 325-340Crossref PubMed Scopus (237) Google Scholar) and localization of ARG with actin microfilaments (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar) have supported a in regulation of the actin The present for involvement of ARG in the cellular response to oxidative stress. Moreover, the of apoptosis by oxidative stress is attenuated in ARG-deficient cells. These findings in to the actin ARG in that an apoptotic with of the family and induces apoptosis in cells Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar). Siva-1 has also been in the of apoptosis A. K. A. R. T. J. PubMed Scopus Google Scholar). Siva-1 but which sequences by induces the apoptotic response Y. Y. Oncogene. PubMed Scopus Google Scholar). The present studies demonstrate that ARG both Siva-1 and the interaction ARG and but is with the of also demonstrate that mutation of the Siva-1 the apoptotic function of Siva-1 and that apoptosis by Siva-1 is on of These findings ARG as an of Siva-1 in the apoptotic response to oxidative stress. the findings in the that ARG associates with the Siva-1 protein from cells expressing ARG and Siva-1 to with of the by with demonstrated the presence of ARG-Siva-1 of from cells expressing by with support for binding of ARG and Siva-1 from cells expressing with a protein. of the with the binding of ARG and Siva-1 studies with proteins from the ARG and ARG domains demonstrated that both binding to Siva-1 binding of SH2, but SH3, to the protein but a for binding to ARG however, the is in Siva-1 Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Y. Y. Oncogene. PubMed Scopus Google Scholar). In this SH2, and SH3, to Siva-1 These findings demonstrate that the ARG with Siva-1 and of both and and that binding of the ARG is also with Siva-1 is a for ARG, with ARG in the presence of of the by and demonstrated phosphorylation of Siva-1 a no phosphorylation Siva-1 with in which in the binding mutated to are tyrosine phosphorylation sites in Siva-1 that are and of sites to and of the proteins with ARG demonstrated of phosphorylation with but with ARG Siva-1 in with ARG and by with The show that Siva-1 is by ARG in cells no tyrosine phosphorylation of this with also by ARG, no tyrosine phosphorylation of In with findings and the presence of in of with ARG, but also in tyrosine phosphorylation These findings support for ARG-mediated phosphorylation of Siva-1 and on and in ARG, c-Abl R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, P. S. Weichselbaum R. Kharbanda S. Kufe D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), is activated by cells expressing with of for phosphorylation of demonstrated of ARG a the to In studies of and of in phosphorylation of the cells with however, with tyrosine phosphorylation of no phosphorylation of in These findings demonstrate that activation of ARG by is with phosphorylation of Siva-1 on Tyr48. the functional significance of the ARG-Siva-1 cells that stably ARG the no of ARG on cell cell of Siva-1 in the cells with the of apoptotic cells DNA apoptosis in of Siva-1 on cells a of in apoptosis in cells as with that with and no on the and cells Siva-1 expressed in the The demonstrate Siva-1 induces apoptosis in of Siva-1 in the of ARG In with the finding that ARG Siva-1 on the of on the of whereas the proapoptotic effects of to with Siva-1 These findings demonstrate that apoptosis is on the ARG kinase function and that ARG-mediated phosphorylation of Siva-1 on is a proapoptotic the findings demonstrate that ARG is activated by apoptosis in the and cells. The demonstrate that the apoptotic effects of are attenuated in as with cells however, a in apoptosis in cells The apoptotic effects of attenuated as with Moreover, of ARG in cells corrected the defect in apoptosis The ARG tyrosine kinase been with the of S. A. M. P. A. PubMed Google Scholar, Y. T. T. M. M. N. K. S. Y. Y. Google Scholar). the to c-Abl and a in neurolation as (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar), a function for ARG in cell has studies have that Abl family kinases cellular with the cytoskeleton Cell. 1989; 58: Full Text PDF PubMed Scopus Google K.K. PubMed Scopus Google Scholar). The of actin-binding domains in the C-terminal region of ARG (3Van Etten R.A. Jackson P.K. Baltimore D. Sanders M.C. Matsuddaira P.T. Janmey P.A. J. Cell Biol. 1994; 124: 325-340Crossref PubMed Scopus (237) Google Scholar) and localization of ARG with actin microfilaments (10Koleske A.J. Gifford A.M. Scott M.L. Nee M. Bronson R.T. Miczek K.A. Baltimore D. Neuron. 1998; 21: 1259-1272Abstract Full Text Full Text PDF PubMed Scopus (333) Google Scholar) have supported a in regulation of the actin The present for involvement of ARG in the cellular response to oxidative stress. Moreover, the of apoptosis by oxidative stress is attenuated in ARG-deficient cells. These findings in to the actin ARG in that an apoptotic Siva-1 with of the family and induces apoptosis in cells Y. M. S. Proc. Natl. Acad. Sci. U. S. A. 1997; PubMed Scopus Google Scholar). Siva-1 has also been in the of apoptosis A. K. A. R. T. J. PubMed Scopus Google Scholar). Siva-1 but which sequences by induces the apoptotic response Y. Y. Oncogene. PubMed Scopus Google Scholar). The present studies demonstrate that ARG both Siva-1 and the interaction ARG and but is with the of also demonstrate that mutation of the Siva-1 the apoptotic function of Siva-1 and that apoptosis by Siva-1 is on of These findings ARG as an of Siva-1 in the apoptotic response to oxidative stress. The ARG tyrosine kinase with Siva-1 in the apoptotic response to oxidative of PDF