JBC INTERFERin siRNA transfection reagent

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M413787200 on January 18, 2005

J. Biol. Chem., Vol. 280, Issue 12, 11147-11151, March 25, 2005
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
280/12/11147    most recent
M413787200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Raina, D.
Right arrow Articles by Kufe, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Raina, D.
Right arrow Articles by Kufe, D.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

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

Deepak Raina{ddagger}, Pramod Pandey{ddagger}§, Rehan Ahmad{ddagger}, Ajit Bharti{ddagger}, Jian Ren{ddagger}, Surender Kharbanda{ddagger}, Ralph Weichselbaum||, and Donald Kufe{ddagger}**

From the {ddagger}Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts 02115 and the ||Department of Radiation and Cellular Oncology, University of Chicago Medical School, Chicago, Illinois 60637

Received for publication, December 7, 2004 , and in revised form, January 10, 2005.


   ABSTRACT
TOP
 ABSTRACT
INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
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.


   INTRODUCTION
TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
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 (1–3). 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 (4–6). 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 (7, 8). 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 (4, 9). 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 (10), which maintains caspase-9 in the inactive monomer conformation (11, 12) and functions as a tether for caspase-3 (13, 14). Other studies have demonstrated that caspase-9 activity is inhibited by Akt-mediated phosphorylation on Ser-196 (15) and by extracellular signal-regulated kinase-mediated phosphorylation on Thr-125 (16). 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 (17). The product of the gene mutated in ataxia telangiectasia is responsible in part for c-Abl activation (18, 19). Other work has demonstrated that nuclear c-Abl interacts with the DNA-dependent protein kinase (DNA-PK)-Ku complex (20, 21). Phosphorylation of c-Abl by the catalytic subunit DNA-PKcs stimulates c-Abl activity (20). 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 (22–26). 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 (17, 27–29). Moreover, c-Abl interactions with Rad51, Rad9, and the hTERT telomerase catalytic subunit have been implicated in the apoptotic response to DNA damage (30–32). 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 (23, 33, 34). 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.


   MATERIALS AND METHODS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
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 (35) were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and antibiotics. Cells were treated with 10 µM 1-({beta}-D-arabinofuranosyl)cytosine (araC)1 (Sigma). Irradiation was performed with a {gamma}-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 (36). 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 (37) and cleared by centrifugation at 12,000 x g for 15 min. Cytosolic S15 lysates were prepared as described (14). 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 (36), anti-caspase-3 (sc-7148; Santa Cruz) and anti-{beta}-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 buffer for 2 h at 4 °C. In other experiments, GST, GST-c-Abl SH3, GST-c-Abl, or GST-c-Abl(K-R) were incubated with purified His-caspase-9. Adsorbates to glutathione beads were analyzed by immunoblotting.

In Vitro Kinase Assays—GST-c-Abl and GST-c-Abl(K-R) purified from baculovirus-infected Sf9 cells (24) were incubated in kinase buffer (50 mM HEPES, pH 7.4, 10 mM MgCl2, 10 mM MnCl2, 2 mM dithiothreitol, 0.1 mM sodium vanadate) with substrate and [{gamma}-32P]ATP (3,000 Ci/mmol; PerkinElmer Life Sciences) for 15 min at 30 °C. Reaction products were analyzed by SDS-PAGE and autoradiography.

Apoptosis Assays—DNA content was assessed by staining ethanol-fixed cells with propidium iodide and monitoring by FACScan (Becton Dickinson). Percentage of cells with sub-G1 DNA were determined by the MODFIT LT Program (Verity Software). DNA laddering was performed as described (38).


   RESULTS
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
c-Abl Associates with Caspase-9—To determine whether c-Abl associates with caspase-9, lysates from U-937 cells were immunoprecipitated with anti-caspase-9. Immunoblot analysis of the precipitates with anti-c-Abl demonstrated little if any signal (Fig. 1A). By contrast, complexes of c-Abl and caspase-9 were detectable when similar experiments were performed on U-937 cells treated with araC, an inhibitor of DNA replication (39) and inducer of c-Abl activation (33, 34) (Fig. 1A). The association of c-Abl and caspase-9 was found at 30 min of araC treatment and was maximal at 60–120 min (Fig. 1A). To confirm the association, U-937 cell lysates were incubated with GST or a GST-caspase-9 fusion protein. Immunoblot analysis of the adsorbates showed binding of c-Abl to GST-caspase-9 and not GST (Fig. 1B). To determine whether the interaction is direct, purified His-tagged caspase-9 was incubated with GST fusion proteins containing kinase-active GST-c-Abl or kinase-inactive GST-c-Abl(K-R). Immunoblotting of the adsorbates with anti-caspase-9 demonstrated binding to both c-Abl and c-Abl(K-R) (Fig. 1C). To further define the c-Abl sequences responsible for binding to caspase-9, we incubated Hiscaspase-9 with GST or a GST fusion protein that contains the c-Abl SH3 domain (GST-c-Abl SH3). The results show that c-Abl SH3 binds to His-caspase-9 (Fig. 1D). These findings indicate that c-Abl associates with caspase-9 in the response to genotoxic stress and that the interaction is mediated by direct binding through the c-Abl SH3 domain.



View larger version (33K):
[in this window]
[in a new window]
  		FIG. 1.
c-Abl associates with caspase-9. A, U-937 cells were left untreated or treated with 10 µM araC for the indicated times. Cell lysates were immunoprecipitated with anti-caspase-9. Immune complexes were subjected to immunoblotting (IB) with anti-c-Abl and anti-caspase-9. B, U-937 cell lysates were incubated with GST or GST-caspase-9. The adsorbates were analyzed by immunoblotting with anti-c-Abl. C, column-purified His-caspase-9 was incubated with glutathione beads containing GST, GST-c-Abl, or GST-c-Abl(K-R). The adsorbates were subjected to immunoblot analysis with anti-caspase-9. D, His-caspase-9 was incubated with GST or GST-c-Abl SH3. The adsorbates were immunoblotted with anti-caspase-9.

 
c-Abl Phosphorylates Caspase-9 on Tyr-153—To determine whether caspase-9 functions as a c-Abl substrate, we incubated His-caspase-9 with purified GST-c-Abl or GST-c-Abl(K-R) and [{gamma}-32P]ATP. Analysis of the products by SDS-PAGE and auto-radiography showed that caspase-9 is phosphorylated by c-Abl and not c-Abl(K-R) (Fig. 2A, left). To define the phosphorylation site, individual tyrosines in His-caspase-9 were mutated to phenylalanine (Tyr -> Phe), and the mutants were analyzed as substrates for c-Abl. The results demonstrated that, in contrast to other mutants (Y251F, Y345F, Y363F, Y397F), there was no detectable phosphorylation of His-caspase-9 with the Y153F mutation (Fig. 2A, right). To determine whether c-Abl phosphorylates caspase-9 in vivo, studies were performed with wild-type and c-abl–/– mouse fibroblasts. Constitutive and araC-induced tyrosine phosphorylation of caspase-9 was detectable in wild-type, but not c-abl–/–, cells (Fig. 2B). Importantly, expression of c-Abl in the c-abl–/– cells (c-abl+/–) was also associated with constitutive and araC-induced tyrosine phosphorylation of caspase-9 (Fig. 2B). To determine whether caspase-9 is phosphorylated on Tyr-153 in cells, we generated U-937 cells stably expressing an empty vector or caspase-9(Y153F) (Fig. 2C, left). U-937 cells transfected with wild-type caspase-9 were not viable. As found for endogenous caspase-9, araC treatment was associated with coprecipitation of c-Abl and caspase-9(Y153F) (Fig. 2C, right). In addition, immunoblot analysis of anti-caspase-9 immunoprecipitates with anti-phospho-Tyr demonstrated araC-induced tyrosine phosphorylation of caspase-9 in the U-937/vector cells (Fig. 2D, left). By contrast, there was little detectable tyrosine phosphorylation of caspase-9 in the araC-treated U-937/caspase-9(Y153F) cells (Fig. 2D, right). These findings indicate that c-Abl phosphorylates caspase-9 on Tyr-153 in vitro and in the response of cells to DNA damage.



View larger version (40K):
[in this window]
[in a new window]
  		FIG. 2.
c-Abl phosphorylates caspase-9 on Tyr-153 in vitro and in vivo in response to DNA damage. A, purified recombinant c-Abl or c-Abl(K-R) was incubated with His-caspase-9 and [{gamma}-32P]ATP (left). His-caspase-9, His-caspase-9(Y153F) were incubated with recombinant c-Abl and [{gamma}-32P]ATP (right). The reaction products were analyzed by SDS-PAGE and autoradiography (upper panels) or Coomassie blue staining (lower panels). B, wild-type, c-abl–/– or c-abl+/– cells were treated with 10 µM araC for indicated times. Anti-caspase-9 immunoprecipitates were analyzed by immunoblotting with anti-phospho-Tyr or anti-caspase-9. C, lysates from U-937/vector or U-937/caspase-9(Y153F) cells (lanes A and B represent two separately isolated clones) were subjected to immunoblot analysis with anti-caspase-9 and anti-{beta}-actin (left). U-937/caspase-9(Y153F) cells were treated with 10 µM araC for 1 h. Cell lysates were immunoprecipitated with anti-caspase-9. Immune complexes were subjected to immunoblotting with anti-c-Abl and anti-caspase-9 (right). D, U-937/vector (left) and U-937/caspase-9(Y153F) (right) cells were treated with 10 µM araC for indicated times. Anti-caspase-9 immunoprecipitates were analyzed by immunoblotting with anti-phospho-Tyr and anti-caspase-9.

 
Caspase-9(Y153F) or STI571 Treatment Attenuates DNA Damage-induced Autoprocessing of Caspase-9—Treatment of U-937 cells with araC is associated with release of mitochondrial cytochrome c to the cytosol and activation of caspase-3 at 4 and 6 h (34). araC treatment of U-937/vector cells was also associated with processing of caspase-9 to the p37 and p35 subunits (Fig. 3A). Importantly, however, processing of caspase-9 to the p35 fragment was attenuated in the araC-treated U-937/caspase-9(Y153F) cells (Fig. 3A). Moreover, araC-induced activation of caspase-3 was attenuated in U-937/caspase-9(Y153F) cells compared with that in U-937/vector cells (Fig. 3B). Caspase-3 induces cleavage of caspase-9 to the p37 subunit (4, 9). Thus, cleavage of caspase-9(Y153F) to p37 in the absence of caspase-3 activation indicates that low levels of caspase-3 activity or other proteases, perhaps even caspase-9 itself, may induce cleavage at Asp-330 in the presence of the Y153F mutation. To determine whether inhibition of c-Abl activity has similar effects on caspase-9 and caspase-3, U-937 cells were treated with the c-Abl inhibitor, STI571. Exposure to STI571 had no apparent effect on araC-induced release of cytochrome c (Fig. 3C). However, STI571 treatment was associated with attenuation of araC-induced cleavage of caspase-9 to the p35 subunit (Fig. 3C). STI571 also attenuated araC-induced activation of caspase-3 (data not shown). Consistent with these results, analysis of anti-caspase-9 immunoprecipitates from cytosolic fractions of U-937 cells demonstrated that araC increases the formation of c-Abl-caspase-9 complexes and that STI571 attenuates this response (Fig. 3D). Moreover, STI571 attenuated the araC-induced cleavage of caspase-9 to p35 (Fig. 3D). These findings indicate that, like expression of caspase-9(Y153F), inhibition of the c-Abl kinase with STI571 attenuates araC-induced autoprocessing of caspase-9 to the p35 subunit.



View larger version (54K):
[in this window]
[in a new window]
  		FIG. 3.
Caspase-9(Y153F) and STI571 attenuate DNA damage-induced autocleavage of caspase-9. A and B, U-937/vector or U-937/caspase-9(Y153F) cells were treated with 10 µM araC for the indicated times. Cytoplasmic fractions were subjected to immunoblotting with anti-caspase-9 and {beta}-actin (left) or with anti-caspase-3 and anti-{beta}-actin (right). C and D, U-937 cells were pretreated with 10 µM STI571 for 24 h and then exposed to 10 µM araC for 4 h. Cytoplasmic fractions were subjected to immunoblot analysis with the indicated antibodies (C). Anti-caspase-9 immunoprecipitates were analyzed by immunoblotting with the indicated antibodies (D).

 
Caspase-9(Y153F) Attenuates DNA Damage-induced Apoptosis—The effects of the caspase-9(Y153F) mutant on araC-induced apoptosis were first assessed by monitoring the percentage of cells with sub-G1 DNA. araC treatment of U-937/vector cells was associated with a substantial increase in sub-G1 DNA content, and this response was attenuated in U-937/caspase-9(Y153F) cells (supplemental Fig. S1A). Similar results were obtained at different times of araC exposure and in repetitive experiments (Fig. 4A). U-937 cells also respond to 20 grays of ionizing radiation (IR) with the induction of apoptosis (40). As found with araC, IR-induced apoptosis of U-937/caspase-9(Y153F) cells was decreased compared with that obtained with U-937/vector cells (supplemental Fig. S1B). Confirmation of these results in additional experiments (Fig. 4B) indicated that U-937/caspase-9(Y153F) cells exhibit an attenuated apoptotic response to diverse DNA damaging agents. As an additional assay for apoptotic cells, araC-induced DNA laddering was attenuated in U-937 cells expressing caspase-9(Y153F) (Fig. 4C). Similar results were obtained with IR-treated U-937/vector and U-937/caspase-9(Y153F) cells (Fig. 4C). These findings indicate that caspase-9(Y153F) attenuates DNA damage-induced apoptosis.



View larger version (26K):
[in this window]
[in a new window]
  		FIG. 4.
Caspase-9(Y153F) attenuates DNA damage-induced apoptosis. A, U-937/vector (open bars) and U-937/caspase-9(Y153F) (solid bars) cells were treated with 10 µM araC for the indicated times. The results are expressed as the percentage (mean ± S.D. of three independent experiments) of apoptotic cells with sub-G1 DNA. B, U-937/vector (open bars) and U-937/caspase-9(Y153F) (solid bars) cells were treated with 20 grays of IR and harvested at the indicated times. The percentage of apoptotic cells with sub-G1 DNA is expressed as the mean ± S.D. of three independent experiments. C, U-937/vector and U-937/caspase-9(Y153F) cells were treated with araC for 18 h or with 20 grays of IR and harvested at 24 h. The cells were then assayed for DNA fragmentation by agarose gel electrophoresis.

 

   DISCUSSION
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
c-Abl Phosphorylates Caspase-9 on Tyr-153—Previous studies have demonstrated that Akt and extracellular signal-regulated kinase phosphorylate and thereby inactivate caspase-9 processing (15, 16). The present work demonstrates that c-Abl associates with caspase-9 in the response of cells to genotoxic stress. Low levels of constitutive binding of c-Abl and caspase-9 were also observed when cells were stressed by being overgrown (data not shown). In vitro studies further demonstrated that the c-Abl SH3 domain binds directly to caspase-9. The available evidence indicates that the c-Abl SH3 domain interacts with a proline-rich motif in the caspase-9 caspase recruitment domain region (PEVLRPETP; amino acids 118–126) (data not shown). The results also demonstrate that c-Abl phosphorylates caspase-9 on Tyr-153. The Tyr-153 site resides in the caspase-9 large subunit near the caspase recruitment domain region. Based on structural studies, Tyr-153 would be exposed on the inactive caspase-9 conformation and possibly buried in the active conformation near the dimer interface (7). Consistent with these predictions, caspase-9 and not caspase-9(Y153F) was subject to tyrosine phosphorylation in the response to DNA damage. DNA damage-induced phosphorylation of caspase-9 by a c-Abl-dependent mechanism was confirmed by the absence of this response in cells null for c-Abl expression. Moreover, expression of c-Abl in c-abl+/– cells was associated with tyrosine phosphorylation of caspase-9. These findings indicate that caspase-9 is phosphorylated by c-Abl on Tyr-153 in vitro and in the cellular response to DNA damage.

c-Abl Regulates Caspase-9 Autoprocessing in the Apoptotic Response to DNA Damage—c-Abl is activated in the response of cells to genotoxic stress (17). Other studies on wild-type, c-abl–/–, and c-abl+/– cells have shown that c-Abl is essential for DNA damage-induced apoptosis (23, 33). Moreover, inhibition of c-Abl with STI571 treatment has been shown to block araC-induced apoptosis (34). c-Abl interacts with diverse signaling molecules that are associated with the apoptotic response (17, 22–32); however, there has been no known interaction between c-Abl and the initiator or effector caspases. In the present studies, the functional significance of the c-Abl-caspase-9 interaction in cells is supported by the demonstration that inhibition of c-Abl with STI571 blocks autoprocessing of caspase-9 to the p35 subunit. Caspase-9(Y153F) also blocked processing to the p35 subunit and caspase-3 activation. Our results further demonstrate that caspase-9(Y153F) blocks araC-induced apoptosis. araC misincorporates into DNA and functions as an inhibitor of DNA replication (39). The demonstration that caspase-9(Y153F) similarly attenuates IR-induced apoptosis indicates that the c-Abl -> caspase-9 pathway is of importance in the response to other genotoxic agents. Of potential significance, caspase-9 Tyr-153 is conserved in sites that are proximal to L1 loops in other caspases, including caspase-3 (Tyr-37) and caspase-7 (Tyr-60). The present findings thus support a model in which caspase-9 autocleavage is regulated by a c-Abl-dependent mechanism in the apoptotic response to genotoxic stress.


   FOOTNOTES
 
* This work was supported by NCI, National Institutes of Health Grant CA29431. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Back

{boxs} The on-line version of this article (available at http://www.jbc.org) contains a supplemental figure. Back

§ Present address: Suntory Pharmaceutical Research Laboratories, Cambridge, MA 02139. Back

** To whom correspondence should be addressed: Dept. of Adult Oncology, Dana-Farber Cancer Inst., 44 Binney St., Boston, MA 02115. Tel.: 617-632-3141; Fax: 617-632-2934; E-mail: donald_kufe{at}dfci.harvard.edu.

1 The abbreviations used are: araC, 1-({beta}-D-arabinofuranosyl)cytosine; GST, glutathione S-transferase; SH3, Src homology 3; IR, ionizing radiation. Back


   ACKNOWLEDGMENTS
 
We thank Kamal Chauhan for technical support.



   REFERENCES
TOP
 ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. Adrain, C., Slee, E., Harte, M., and Martin, S. (1999) J. Biol. Chem. 274, 20855–20860[Abstract/Free Full Text]
  2. Zou, H., Li, X., Liu, X., and Wang, X. (1999) J. Biol. Chem. 274, 11549–11556[Abstract/Free Full Text]
  3. Jiang, X., and Wang, X. (2000) J. Biol. Chem. 275, 31199–31203[Abstract/Free Full Text]
  4. Srinivasula, S., Ahmad, M., Alnemri, T., and Alnemri, E. (1998) Mol. Cell 1, 949–957[CrossRef][Medline] [Order article via Infotrieve]
  5. Stennicke, H., Deveraux, Q., Humke, E., Reed, J., Dixit, V., and Salvesen, G. (1999) J. Biol. Chem. 26, 8359–8362
  6. Rodriguez, J., and Lazebnik, Y. (1999) Genes Dev. 13, 3179–3184[Abstract/Free Full Text]
  7. Renatus, M., Stennicke, H. R., Scott, F. L., Liddington, R. C., and Salvesen, G. S. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 14250–14255[Abstract/Free Full Text]
  8. Shiozaki, E. N., Chai, J., and Shi, Y. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 4197–4202[Abstract/Free Full Text]
  9. Slee, 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., and Martin, S. J. (1999) J. Cell Biol. 144, 281–292[Abstract/Free Full Text]
  10. Srinivasula, S. M., Hegde, R., Saleh, A., Datta, P., Shiozaki, E., Chai, J., Lee, R. A., Robbins, P. D., Fernandes-Alnemri, T., Shi, Y., and Alnemri, E. S. (2001) Nature 410, 112–116[CrossRef][Medline] [Order article via Infotrieve]
  11. Fesik, S. W., and Shi, Y. (2001) Science 294, 1477–1478[Free Full Text]
  12. Shiozaki, E. N., Chai, J., Rigotti, D. J., Riedl, S. J., Li, P., Srinivasula, S. M., Alnemri, E. S., Fairman, R., and Shi, Y. (2003) Mol. Cell 11, 519–527[CrossRef][Medline] [Order article via Infotrieve]
  13. Bratton, S. B., Walker, G., Srinivasula, S. M., Sun, X. M., Butterworth, M., Alnemri, E. S., and Cohen, G. M. (2001) EMBO J. 20, 998–1009[CrossRef][Medline] [Order article via Infotrieve]
  14. Hill, M. M., Adrain, C., Duriez, P. J., Creagh, E. M., and Martin, S. J. (2004) EMBO J. 23, 2134–2145[CrossRef][Medline] [Order article via Infotrieve]
  15. Cardone, M., Roy, N., Stennicke, H., Salvesen, G., Franke, T., Stanbridge, E., Frisch, S., and Reed, J. (1998) Science 282, 1318–1321[Abstract/Free Full Text]
  16. Allan, L. A., Morrice, N., Brady, S., Magee, G., Pathak, S., and Clarke, P. R. (2003) Nat. Cell Biol. 5, 647–654[CrossRef][Medline] [Order article via Infotrieve]
  17. Kharbanda, S., Ren, R., Pandey, P., Shafman, T. D., Feller, S. M., Weichselbaum, R. R., and Kufe, D. W. (1995) Nature 376, 785–788[CrossRef][Medline] [Order article via Infotrieve]
  18. Shafman, 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., and Lavin, M. F. (1997) Nature 387, 520–523[CrossRef][Medline] [Order article via Infotrieve]
  19. Baskaran, 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., and Wang, J. Y. J. (1997) Nature 387, 516–519[CrossRef][Medline] [Order article via Infotrieve]
  20. Kharbanda, S., Pandey, P., Jin, S., Inoue, S., Bharti, A., Yuan, Z.-M., Weichselbaum, R., Weaver, D., and Kufe, D. (1997) Nature 386, 732–735[CrossRef][Medline] [Order article via Infotrieve]
  21. Jin, S., Kharbanda, S., Mayer, B., Kufe, D., and Weaver, D. T. (1997) J. Biol. Chem. 272, 24763–24766[Abstract/Free Full Text]
  22. Yuan, Z., Huang, Y., Fan, M., Sawers, C., Kharbanda, S., and Kufe, D. (1996) J. Biol. Chem. 271, 26457–26460[Abstract/Free Full Text]
  23. Yuan, Z., Huang, Y., Ishiko, T., Kharbanda, S., Weichselbaum, R., and Kufe, D. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 1437–1440[Abstract/Free Full Text]
  24. Yuan, Z. M., Shioya, H., Ishiko, T., Sun, X., Huang, Y., Lu, H., Kharbanda, S., Weichselbaum, R., and Kufe, D. (1999) Nature 399, 814–817[CrossRef][Medline] [Order article via Infotrieve]
  25. Agami, R., Blandino, G., Oren, M., and Shaul, Y. (1999) Nature 399, 809–813[CrossRef][Medline] [Order article via Infotrieve]
  26. Gong, J., Costanzo, A., Yang, H., Melino, G., Kaelin, W., Jr., Levrero, M., and Wang, J. Y. J. (1999) Nature 399, 806–809[CrossRef][Medline] [Order article via Infotrieve]
  27. Kharbanda, S., Pandey, P., Ren, R., Feller, S., Mayer, B., Zon, L., and Kufe, D. (1995) J. Biol. Chem. 270, 30278–30281[Abstract/Free Full Text]
  28. Pandey, P., Raingeaud, J., Kaneki, M., Weichselbaum, R., Davis, R., Kufe, D., and Kharbanda, S. (1996) J. Biol. Chem. 271, 23775–23779[Abstract/Free Full Text]
  29. Kharbanda, S., Pandey, P., Yamauchi, T., Kumar, S., Kaneki, M., Kumar, V., Bharti, A., Yuan, Z., Ghanem, L., Rana, A., Weichselbaum, R., Johnson, G., and Kufe, D. (2000) Mol. Cell. Biol. 20, 4979–4989[Abstract/Free Full Text]
  30. Yuan, Z. M., Huang, Y., Ishiko, T., Nakada, S., Utsugisawa, T., Kharbanda, S., Sung, P., Shinohara, A., Weichselbaum, R., and Kufe, D. (1998) J. Biol. Chem. 273, 3799–3802[Abstract/Free Full Text]
  31. Kharbanda, S., Kumar, V., Dhar, S., Pandey, P., Chen, C., Majumder, P., Yuan, Z., Whang, Y., Strauss, W., Pandita, T., Weaver, D., and Kufe, D. (2000) Curr. Biol. 10, 568–575[CrossRef][Medline] [Order article via Infotrieve]
  32. Yoshida, K., Komatsu, K., Wang, H.-G., and Kufe, D. (2002) Mol. Cell. Biol. 22, 3292–3300[Abstract/Free Full Text]
  33. Huang, Y., Yuan, Z. M., Ishiko, T., Nakada, S., Utsugisawa, T., Kato, T., Kharbanda, S., and Kufe, D. W. (1997) Oncogene 15, 1947–1952[CrossRef][Medline] [Order article via Infotrieve]
  34. Raina, D., Mishra, N., Kumar, S., Kharbanda, S., Saxena, S., and Kufe, D. (2002) Mol. Pharmacol. 61, 1489–1495[Abstract/Free Full Text]
  35. Tybulewicz, V. L. J., Crawford, C. E., Jackson, P. K., Bronson, R. T., and Mulligan, R. C. (1991) Cell 65, 1153–1163[CrossRef][Medline] [Order article via Infotrieve]
  36. Pandey, P., Saleh, A., Nakazawa, A., Kumar, S., Srinivasula, S. M., Kumar, V., Weichselbaum, R., Nalin, C., Alnemri, E. S., Kufe, D., and Kharbanda, S. (2000) EMBO J. 19, 4310–4322[CrossRef][Medline] [Order article via Infotrieve]
  37. Yoshida, K., Kharbanda, S., and Kufe, D. (1999) J. Biol. Chem. 274, 34663–34668[Abstract/Free Full Text]
  38. Datta, R., Kojima, H., Yoshida, K., and Kufe, D. (1997) J. Biol. Chem. 272, 20317–20320[Abstract/Free Full Text]
  39. Major, P. P., Egan, E. M., Beardsley, G., Minden, M., and Kufe, D. W. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 3235–3239[Abstract/Free Full Text]
  40. Kharbanda, S., Saxena, S., Yoshida, K., Pandey, P., Kaneki, M., Wang, Q., Cheng, K., Chen, Y., Campbell, A., Thangrila, S., Yuan, Z., Narula, J., Weichselbaum, R., Nalin, C., and Kufe, D. (2000) J. Biol. Chem. 275, 322–327[Abstract/Free Full Text]

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
M. A. McDonnell, Md. J. Abedin, M. Melendez, T. N. Platikanova, J. R. Ecklund, K. Ahmed, and A. Kelekar
Phosphorylation of Murine Caspase-9 by the Protein Kinase Casein Kinase 2 Regulates Its Cleavage by Caspase-8
J. Biol. Chem., July 18, 2008; 283(29): 20149 - 20158.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
R. Hagerkvist, S. Sandler, D. Mokhtari, and N. Welsh
Amelioration of diabetes by imatinib mesylate (Gleevec(R)): role of {beta}-cell NF-{kappa}B activation and anti-apoptotic preconditioning
FASEB J, February 1, 2007; 21(2): 618 - 628.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
280/12/11147    most recent
M413787200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Raina, D.
Right arrow Articles by Kufe, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Raina, D.
Right arrow Articles by Kufe, D.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2005 by the American Society for Biochemistry and Molecular Biology.