Caspase-3-dependent Cleavage of Bcl-2 Promotes Release of Cytochrome c
Caspases are cysteine proteases that mediate apoptosis by proteolysis of specific substrates. Although many caspase substrates have been identified, for most substrates the physiologic caspase(s) required for cleavage is unknown. The Bcl-2 protein, which inhibits apoptosis, is cleaved at Asp-34 by c...
Gespeichert in:
Veröffentlicht in: | The Journal of biological chemistry 1999-07, Vol.274 (30), p.21155-21161 |
---|---|
Hauptverfasser: | , , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 21161 |
---|---|
container_issue | 30 |
container_start_page | 21155 |
container_title | The Journal of biological chemistry |
container_volume | 274 |
creator | Kirsch, David G. Doseff, Andrea Chau, B. Nelson Lim, Dae-Sik de Souza-Pinto, Nadja C. Hansford, Richard Kastan, Michael B. Lazebnik, Yuri A. Hardwick, J. Marie |
description | Caspases are cysteine proteases that mediate apoptosis by proteolysis of specific substrates. Although many caspase substrates have been identified, for most substrates the physiologic caspase(s) required for cleavage is unknown. The Bcl-2 protein, which inhibits apoptosis, is cleaved at Asp-34 by caspases during apoptosis and by recombinant caspase-3 in vitro. In the present study, we show that endogenous caspase-3 is a physiologic caspase for Bcl-2. Apoptotic extracts from 293 cells cleave Bcl-2 but not Bax, even though Bax is cleaved to an 18-kDa fragment in SK-NSH cells treated with ionizing radiation. In contrast to Bcl-2, cleavage of Bax was only partially blocked by caspase inhibitors. Inhibitor profiles indicate that Bax may be cleaved by more than one type of noncaspase protease. Immunodepletion of caspase-3 from 293 extracts abolished cleavage of Bcl-2 and caspase-7, whereas immunodepletion of caspase-7 had no effect on Bcl-2 cleavage. Furthermore, MCF-7 cells, which lack caspase-3 expression, do not cleave Bcl-2 following staurosporine-induced cell death. However, transient transfection of caspase-3 into MCF-7 cells restores Bcl-2 cleavage after staurosporine treatment. These results demonstrate that in these models of apoptosis, specific cleavage of Bcl-2 requires activation of caspase-3. When the pro-apoptotic caspase cleavage fragment of Bcl-2 is transfected into baby hamster kidney cells, it localizes to mitochondria and causes the release of cytochrome c into the cytosol. Therefore, caspase-3-dependent cleavage of Bcl-2 appears to promote further caspase activation as part of a positive feedback loop for executing the cell. |
doi_str_mv | 10.1074/jbc.274.30.21155 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_69897350</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0021925819725587</els_id><sourcerecordid>69897350</sourcerecordid><originalsourceid>FETCH-LOGICAL-c464t-e536c506091d603d65f078dfdcfdca87858156e4e1b6297c209ab27b629f756b3</originalsourceid><addsrcrecordid>eNp1kM9LwzAUx4Mobk7vnqQH8db50jRp402LU0FQRMFbSJPXraNdZ9NN9t-brTuI4CPwCN8fPD6EnFMYU0ji63luxlESjxmMI0o5PyBDCikLGaefh2QIENFQRjwdkBPn5uAnlvSYDCjEIIWQQzLJtFtqhyELLS5xYXHRBVmFeq2nGDRFcGeqMApe26ZuOnTBG3rN7ZRs0zVm5gUMzCk5KnTl8Gy_R-Rjcv-ePYbPLw9P2e1zaGIRdyFyJgwHAZJaAcwKXkCS2sIa_3SapDylXGCMNBeRTEwEUudRsv0UCRc5G5GrvnfZNl8rdJ2qS2ewqvQCm5VTQqYyYRy8EXqjaRvnWizUsi1r3W4UBbVlpzw75dkpBmrHzkcu9t2rvEb7K9DD8obL3jArp7PvskWVl54A1n97bnobehDrElvlTIkLg9ZHTKdsU_5_xA-xu4em</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>69897350</pqid></control><display><type>article</type><title>Caspase-3-dependent Cleavage of Bcl-2 Promotes Release of Cytochrome c</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>Kirsch, David G. ; Doseff, Andrea ; Chau, B. Nelson ; Lim, Dae-Sik ; de Souza-Pinto, Nadja C. ; Hansford, Richard ; Kastan, Michael B. ; Lazebnik, Yuri A. ; Hardwick, J. Marie</creator><creatorcontrib>Kirsch, David G. ; Doseff, Andrea ; Chau, B. Nelson ; Lim, Dae-Sik ; de Souza-Pinto, Nadja C. ; Hansford, Richard ; Kastan, Michael B. ; Lazebnik, Yuri A. ; Hardwick, J. Marie</creatorcontrib><description>Caspases are cysteine proteases that mediate apoptosis by proteolysis of specific substrates. Although many caspase substrates have been identified, for most substrates the physiologic caspase(s) required for cleavage is unknown. The Bcl-2 protein, which inhibits apoptosis, is cleaved at Asp-34 by caspases during apoptosis and by recombinant caspase-3 in vitro. In the present study, we show that endogenous caspase-3 is a physiologic caspase for Bcl-2. Apoptotic extracts from 293 cells cleave Bcl-2 but not Bax, even though Bax is cleaved to an 18-kDa fragment in SK-NSH cells treated with ionizing radiation. In contrast to Bcl-2, cleavage of Bax was only partially blocked by caspase inhibitors. Inhibitor profiles indicate that Bax may be cleaved by more than one type of noncaspase protease. Immunodepletion of caspase-3 from 293 extracts abolished cleavage of Bcl-2 and caspase-7, whereas immunodepletion of caspase-7 had no effect on Bcl-2 cleavage. Furthermore, MCF-7 cells, which lack caspase-3 expression, do not cleave Bcl-2 following staurosporine-induced cell death. However, transient transfection of caspase-3 into MCF-7 cells restores Bcl-2 cleavage after staurosporine treatment. These results demonstrate that in these models of apoptosis, specific cleavage of Bcl-2 requires activation of caspase-3. When the pro-apoptotic caspase cleavage fragment of Bcl-2 is transfected into baby hamster kidney cells, it localizes to mitochondria and causes the release of cytochrome c into the cytosol. Therefore, caspase-3-dependent cleavage of Bcl-2 appears to promote further caspase activation as part of a positive feedback loop for executing the cell.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.274.30.21155</identifier><identifier>PMID: 10409669</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Apoptosis ; Caspase 3 ; Caspases - metabolism ; Cricetinae ; Cytochrome c Group - metabolism ; Enzyme Activation ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 - metabolism ; Substrate Specificity</subject><ispartof>The Journal of biological chemistry, 1999-07, Vol.274 (30), p.21155-21161</ispartof><rights>1999 © 1999 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c464t-e536c506091d603d65f078dfdcfdca87858156e4e1b6297c209ab27b629f756b3</citedby><cites>FETCH-LOGICAL-c464t-e536c506091d603d65f078dfdcfdca87858156e4e1b6297c209ab27b629f756b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10409669$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kirsch, David G.</creatorcontrib><creatorcontrib>Doseff, Andrea</creatorcontrib><creatorcontrib>Chau, B. Nelson</creatorcontrib><creatorcontrib>Lim, Dae-Sik</creatorcontrib><creatorcontrib>de Souza-Pinto, Nadja C.</creatorcontrib><creatorcontrib>Hansford, Richard</creatorcontrib><creatorcontrib>Kastan, Michael B.</creatorcontrib><creatorcontrib>Lazebnik, Yuri A.</creatorcontrib><creatorcontrib>Hardwick, J. Marie</creatorcontrib><title>Caspase-3-dependent Cleavage of Bcl-2 Promotes Release of Cytochrome c</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Caspases are cysteine proteases that mediate apoptosis by proteolysis of specific substrates. Although many caspase substrates have been identified, for most substrates the physiologic caspase(s) required for cleavage is unknown. The Bcl-2 protein, which inhibits apoptosis, is cleaved at Asp-34 by caspases during apoptosis and by recombinant caspase-3 in vitro. In the present study, we show that endogenous caspase-3 is a physiologic caspase for Bcl-2. Apoptotic extracts from 293 cells cleave Bcl-2 but not Bax, even though Bax is cleaved to an 18-kDa fragment in SK-NSH cells treated with ionizing radiation. In contrast to Bcl-2, cleavage of Bax was only partially blocked by caspase inhibitors. Inhibitor profiles indicate that Bax may be cleaved by more than one type of noncaspase protease. Immunodepletion of caspase-3 from 293 extracts abolished cleavage of Bcl-2 and caspase-7, whereas immunodepletion of caspase-7 had no effect on Bcl-2 cleavage. Furthermore, MCF-7 cells, which lack caspase-3 expression, do not cleave Bcl-2 following staurosporine-induced cell death. However, transient transfection of caspase-3 into MCF-7 cells restores Bcl-2 cleavage after staurosporine treatment. These results demonstrate that in these models of apoptosis, specific cleavage of Bcl-2 requires activation of caspase-3. When the pro-apoptotic caspase cleavage fragment of Bcl-2 is transfected into baby hamster kidney cells, it localizes to mitochondria and causes the release of cytochrome c into the cytosol. Therefore, caspase-3-dependent cleavage of Bcl-2 appears to promote further caspase activation as part of a positive feedback loop for executing the cell.</description><subject>Animals</subject><subject>Apoptosis</subject><subject>Caspase 3</subject><subject>Caspases - metabolism</subject><subject>Cricetinae</subject><subject>Cytochrome c Group - metabolism</subject><subject>Enzyme Activation</subject><subject>HL-60 Cells</subject><subject>Humans</subject><subject>Proto-Oncogene Proteins c-bcl-2 - metabolism</subject><subject>Substrate Specificity</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM9LwzAUx4Mobk7vnqQH8db50jRp402LU0FQRMFbSJPXraNdZ9NN9t-brTuI4CPwCN8fPD6EnFMYU0ji63luxlESjxmMI0o5PyBDCikLGaefh2QIENFQRjwdkBPn5uAnlvSYDCjEIIWQQzLJtFtqhyELLS5xYXHRBVmFeq2nGDRFcGeqMApe26ZuOnTBG3rN7ZRs0zVm5gUMzCk5KnTl8Gy_R-Rjcv-ePYbPLw9P2e1zaGIRdyFyJgwHAZJaAcwKXkCS2sIa_3SapDylXGCMNBeRTEwEUudRsv0UCRc5G5GrvnfZNl8rdJ2qS2ewqvQCm5VTQqYyYRy8EXqjaRvnWizUsi1r3W4UBbVlpzw75dkpBmrHzkcu9t2rvEb7K9DD8obL3jArp7PvskWVl54A1n97bnobehDrElvlTIkLg9ZHTKdsU_5_xA-xu4em</recordid><startdate>19990723</startdate><enddate>19990723</enddate><creator>Kirsch, David G.</creator><creator>Doseff, Andrea</creator><creator>Chau, B. Nelson</creator><creator>Lim, Dae-Sik</creator><creator>de Souza-Pinto, Nadja C.</creator><creator>Hansford, Richard</creator><creator>Kastan, Michael B.</creator><creator>Lazebnik, Yuri A.</creator><creator>Hardwick, J. Marie</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19990723</creationdate><title>Caspase-3-dependent Cleavage of Bcl-2 Promotes Release of Cytochrome c</title><author>Kirsch, David G. ; Doseff, Andrea ; Chau, B. Nelson ; Lim, Dae-Sik ; de Souza-Pinto, Nadja C. ; Hansford, Richard ; Kastan, Michael B. ; Lazebnik, Yuri A. ; Hardwick, J. Marie</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c464t-e536c506091d603d65f078dfdcfdca87858156e4e1b6297c209ab27b629f756b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1999</creationdate><topic>Animals</topic><topic>Apoptosis</topic><topic>Caspase 3</topic><topic>Caspases - metabolism</topic><topic>Cricetinae</topic><topic>Cytochrome c Group - metabolism</topic><topic>Enzyme Activation</topic><topic>HL-60 Cells</topic><topic>Humans</topic><topic>Proto-Oncogene Proteins c-bcl-2 - metabolism</topic><topic>Substrate Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kirsch, David G.</creatorcontrib><creatorcontrib>Doseff, Andrea</creatorcontrib><creatorcontrib>Chau, B. Nelson</creatorcontrib><creatorcontrib>Lim, Dae-Sik</creatorcontrib><creatorcontrib>de Souza-Pinto, Nadja C.</creatorcontrib><creatorcontrib>Hansford, Richard</creatorcontrib><creatorcontrib>Kastan, Michael B.</creatorcontrib><creatorcontrib>Lazebnik, Yuri A.</creatorcontrib><creatorcontrib>Hardwick, J. Marie</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kirsch, David G.</au><au>Doseff, Andrea</au><au>Chau, B. Nelson</au><au>Lim, Dae-Sik</au><au>de Souza-Pinto, Nadja C.</au><au>Hansford, Richard</au><au>Kastan, Michael B.</au><au>Lazebnik, Yuri A.</au><au>Hardwick, J. Marie</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Caspase-3-dependent Cleavage of Bcl-2 Promotes Release of Cytochrome c</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1999-07-23</date><risdate>1999</risdate><volume>274</volume><issue>30</issue><spage>21155</spage><epage>21161</epage><pages>21155-21161</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Caspases are cysteine proteases that mediate apoptosis by proteolysis of specific substrates. Although many caspase substrates have been identified, for most substrates the physiologic caspase(s) required for cleavage is unknown. The Bcl-2 protein, which inhibits apoptosis, is cleaved at Asp-34 by caspases during apoptosis and by recombinant caspase-3 in vitro. In the present study, we show that endogenous caspase-3 is a physiologic caspase for Bcl-2. Apoptotic extracts from 293 cells cleave Bcl-2 but not Bax, even though Bax is cleaved to an 18-kDa fragment in SK-NSH cells treated with ionizing radiation. In contrast to Bcl-2, cleavage of Bax was only partially blocked by caspase inhibitors. Inhibitor profiles indicate that Bax may be cleaved by more than one type of noncaspase protease. Immunodepletion of caspase-3 from 293 extracts abolished cleavage of Bcl-2 and caspase-7, whereas immunodepletion of caspase-7 had no effect on Bcl-2 cleavage. Furthermore, MCF-7 cells, which lack caspase-3 expression, do not cleave Bcl-2 following staurosporine-induced cell death. However, transient transfection of caspase-3 into MCF-7 cells restores Bcl-2 cleavage after staurosporine treatment. These results demonstrate that in these models of apoptosis, specific cleavage of Bcl-2 requires activation of caspase-3. When the pro-apoptotic caspase cleavage fragment of Bcl-2 is transfected into baby hamster kidney cells, it localizes to mitochondria and causes the release of cytochrome c into the cytosol. Therefore, caspase-3-dependent cleavage of Bcl-2 appears to promote further caspase activation as part of a positive feedback loop for executing the cell.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>10409669</pmid><doi>10.1074/jbc.274.30.21155</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0021-9258 |
ispartof | The Journal of biological chemistry, 1999-07, Vol.274 (30), p.21155-21161 |
issn | 0021-9258 1083-351X |
language | eng |
recordid | cdi_proquest_miscellaneous_69897350 |
source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
subjects | Animals Apoptosis Caspase 3 Caspases - metabolism Cricetinae Cytochrome c Group - metabolism Enzyme Activation HL-60 Cells Humans Proto-Oncogene Proteins c-bcl-2 - metabolism Substrate Specificity |
title | Caspase-3-dependent Cleavage of Bcl-2 Promotes Release of Cytochrome c |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T02%3A47%3A58IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Caspase-3-dependent%20Cleavage%20of%20Bcl-2%20Promotes%20Release%20of%20Cytochrome%20c&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Kirsch,%20David%20G.&rft.date=1999-07-23&rft.volume=274&rft.issue=30&rft.spage=21155&rft.epage=21161&rft.pages=21155-21161&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.274.30.21155&rft_dat=%3Cproquest_cross%3E69897350%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=69897350&rft_id=info:pmid/10409669&rft_els_id=S0021925819725587&rfr_iscdi=true |