Identification of a Novel Phosphorylation Site, Ser-170, as a Regulator of Bad Pro-apoptotic Activity
Bad is a pro-apoptotic member of the Bcl-2 family of proteins that is thought to exert a death-promoting effect by heterodimerization with Bcl-XL, nullifying its anti-apoptotic activity. Growth factors may promote cell survival at least partially through phosphorylation of Bad at one or more of Ser-...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 2002-02, Vol.277 (8), p.6399-6405 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 6405 |
|---|---|
| container_issue | 8 |
| container_start_page | 6399 |
| container_title | The Journal of biological chemistry |
| container_volume | 277 |
| creator | Dramsi, Shaynoor Scheid, Michael P. Maiti, Arpita Hojabrpour, Payman Chen, Xianming Schubert, Kathryn Goodlett, David R. Aebersold, Ruedi Duronio, Vincent |
| description | Bad is a pro-apoptotic member of the Bcl-2 family of proteins that is thought to exert a death-promoting effect by heterodimerization with Bcl-XL, nullifying its anti-apoptotic activity. Growth factors may promote cell survival at least partially through phosphorylation of Bad at one or more of Ser-112, -136, or -155. Our previous work showed that Bad is also phosphorylated in response to cytokines at another site, which we now identify as Ser-170. The functional role of this novel phosphorylation site was assessed by site-directed mutagenesis and analysis of the pro-apoptotic function of Bad in transiently transfected HEK293 and COS-7 cells or by stable expression in the cytokine-dependent cell line, MC/9. In general, mutation of Ser-170 to Ala results in a protein with increased ability to induce apoptosis, similar to the S112A mutant. Mutation of Ser-170 to Asp, mimicking a constitutively phosphorylated site, results in a protein that is virtually unable to induce apoptosis. Similarly, the S112A/S170D double mutant does not cause apoptosis in HEK293 and MC/9 cell lines. These data strongly suggest that phosphorylation of Bad at Ser-170 is a critical event in blocking the pro-apoptotic activity of Bad. |
| doi_str_mv | 10.1074/jbc.M109990200 |
| format | Article |
| fullrecord | <record><control><sourceid>hal_cross</sourceid><recordid>TN_cdi_hal_primary_oai_HAL_pasteur_03262538v1</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0021925819824201</els_id><sourcerecordid>oai_HAL_pasteur_03262538v1</sourcerecordid><originalsourceid>FETCH-LOGICAL-c445t-a355f63dc86b9b2e22aab525d534b80f14ea142dbfda482c0c7da2427e7b5ac13</originalsourceid><addsrcrecordid>eNp1kE1vEzEQQC1ERUPhyhGtOHeDP-PdY6iAVgpt1YLEzRrbs11XSbyynaD8ezbaiJ7wZQ7znjV6hHxgdM6olp-frZv_YLRtW8opfUVmjDaiFor9fk1mlHJWt1w15-Rtzs90fLJlb8g5Y5ppQdsZwRuP2xK64KCEuK1iV0F1G_e4ru77mIc-psN6Wj2GgpfVI6aaaXpZQR7JB3zajeuYjuIX8NV9ijUMcSixBFctXQn7UA7vyFkH64zvT_OC_Pr29efVdb26-35ztVzVTkpVahBKdQvhXbOwreXIOYBVXHklpG1oxyQCk9zbzoNsuKNOe-CSa9RWgWPigtTTvz2szZDCBtLBRAjmerkyA-SCu2So4AuuRLM_8vOJdynmnLD7JzFqjn3N2Ne89B2Fj5Mw7OwG_Qt-CjoCn04XhKf-T0hobIiux43hWpvGLER7hJoJwrHFPmAy2QXcOvSj4IrxMfzvgL-VQZQB</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype></control><display><type>article</type><title>Identification of a Novel Phosphorylation Site, Ser-170, as a Regulator of Bad Pro-apoptotic Activity</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>Dramsi, Shaynoor ; Scheid, Michael P. ; Maiti, Arpita ; Hojabrpour, Payman ; Chen, Xianming ; Schubert, Kathryn ; Goodlett, David R. ; Aebersold, Ruedi ; Duronio, Vincent</creator><creatorcontrib>Dramsi, Shaynoor ; Scheid, Michael P. ; Maiti, Arpita ; Hojabrpour, Payman ; Chen, Xianming ; Schubert, Kathryn ; Goodlett, David R. ; Aebersold, Ruedi ; Duronio, Vincent</creatorcontrib><description>Bad is a pro-apoptotic member of the Bcl-2 family of proteins that is thought to exert a death-promoting effect by heterodimerization with Bcl-XL, nullifying its anti-apoptotic activity. Growth factors may promote cell survival at least partially through phosphorylation of Bad at one or more of Ser-112, -136, or -155. Our previous work showed that Bad is also phosphorylated in response to cytokines at another site, which we now identify as Ser-170. The functional role of this novel phosphorylation site was assessed by site-directed mutagenesis and analysis of the pro-apoptotic function of Bad in transiently transfected HEK293 and COS-7 cells or by stable expression in the cytokine-dependent cell line, MC/9. In general, mutation of Ser-170 to Ala results in a protein with increased ability to induce apoptosis, similar to the S112A mutant. Mutation of Ser-170 to Asp, mimicking a constitutively phosphorylated site, results in a protein that is virtually unable to induce apoptosis. Similarly, the S112A/S170D double mutant does not cause apoptosis in HEK293 and MC/9 cell lines. These data strongly suggest that phosphorylation of Bad at Ser-170 is a critical event in blocking the pro-apoptotic activity of Bad.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M109990200</identifier><identifier>PMID: 11717309</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Amino Acid Substitution ; Animals ; Apoptosis ; Apoptosis - physiology ; bcl-Associated Death Protein ; bcl-X Protein ; Carrier Proteins ; Carrier Proteins - chemistry ; Carrier Proteins - metabolism ; Chlorocebus aethiops ; Cloning, Molecular ; COS Cells ; Escherichia coli ; Escherichia coli - genetics ; Genes, bcl-2 ; Humans ; Life Sciences ; Mice ; Mutagenesis, Site-Directed ; Phosphorylation ; Phosphoserine ; Phosphoserine - metabolism ; Poly(ADP-ribose) Polymerases ; Poly(ADP-ribose) Polymerases - metabolism ; Proto-Oncogene Proteins c-bcl-2 ; Proto-Oncogene Proteins c-bcl-2 - metabolism ; Recombinant Proteins ; Recombinant Proteins - chemistry ; Recombinant Proteins - metabolism ; Serine ; Transfection</subject><ispartof>The Journal of biological chemistry, 2002-02, Vol.277 (8), p.6399-6405</ispartof><rights>2002 © 2002 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><rights>Attribution</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c445t-a355f63dc86b9b2e22aab525d534b80f14ea142dbfda482c0c7da2427e7b5ac13</citedby><cites>FETCH-LOGICAL-c445t-a355f63dc86b9b2e22aab525d534b80f14ea142dbfda482c0c7da2427e7b5ac13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,315,781,785,886,27929,27930</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11717309$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://pasteur.hal.science/pasteur-03262538$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Dramsi, Shaynoor</creatorcontrib><creatorcontrib>Scheid, Michael P.</creatorcontrib><creatorcontrib>Maiti, Arpita</creatorcontrib><creatorcontrib>Hojabrpour, Payman</creatorcontrib><creatorcontrib>Chen, Xianming</creatorcontrib><creatorcontrib>Schubert, Kathryn</creatorcontrib><creatorcontrib>Goodlett, David R.</creatorcontrib><creatorcontrib>Aebersold, Ruedi</creatorcontrib><creatorcontrib>Duronio, Vincent</creatorcontrib><title>Identification of a Novel Phosphorylation Site, Ser-170, as a Regulator of Bad Pro-apoptotic Activity</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Bad is a pro-apoptotic member of the Bcl-2 family of proteins that is thought to exert a death-promoting effect by heterodimerization with Bcl-XL, nullifying its anti-apoptotic activity. Growth factors may promote cell survival at least partially through phosphorylation of Bad at one or more of Ser-112, -136, or -155. Our previous work showed that Bad is also phosphorylated in response to cytokines at another site, which we now identify as Ser-170. The functional role of this novel phosphorylation site was assessed by site-directed mutagenesis and analysis of the pro-apoptotic function of Bad in transiently transfected HEK293 and COS-7 cells or by stable expression in the cytokine-dependent cell line, MC/9. In general, mutation of Ser-170 to Ala results in a protein with increased ability to induce apoptosis, similar to the S112A mutant. Mutation of Ser-170 to Asp, mimicking a constitutively phosphorylated site, results in a protein that is virtually unable to induce apoptosis. Similarly, the S112A/S170D double mutant does not cause apoptosis in HEK293 and MC/9 cell lines. These data strongly suggest that phosphorylation of Bad at Ser-170 is a critical event in blocking the pro-apoptotic activity of Bad.</description><subject>Amino Acid Substitution</subject><subject>Animals</subject><subject>Apoptosis</subject><subject>Apoptosis - physiology</subject><subject>bcl-Associated Death Protein</subject><subject>bcl-X Protein</subject><subject>Carrier Proteins</subject><subject>Carrier Proteins - chemistry</subject><subject>Carrier Proteins - metabolism</subject><subject>Chlorocebus aethiops</subject><subject>Cloning, Molecular</subject><subject>COS Cells</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Genes, bcl-2</subject><subject>Humans</subject><subject>Life Sciences</subject><subject>Mice</subject><subject>Mutagenesis, Site-Directed</subject><subject>Phosphorylation</subject><subject>Phosphoserine</subject><subject>Phosphoserine - metabolism</subject><subject>Poly(ADP-ribose) Polymerases</subject><subject>Poly(ADP-ribose) Polymerases - metabolism</subject><subject>Proto-Oncogene Proteins c-bcl-2</subject><subject>Proto-Oncogene Proteins c-bcl-2 - metabolism</subject><subject>Recombinant Proteins</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - metabolism</subject><subject>Serine</subject><subject>Transfection</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kE1vEzEQQC1ERUPhyhGtOHeDP-PdY6iAVgpt1YLEzRrbs11XSbyynaD8ezbaiJ7wZQ7znjV6hHxgdM6olp-frZv_YLRtW8opfUVmjDaiFor9fk1mlHJWt1w15-Rtzs90fLJlb8g5Y5ppQdsZwRuP2xK64KCEuK1iV0F1G_e4ru77mIc-psN6Wj2GgpfVI6aaaXpZQR7JB3zajeuYjuIX8NV9ijUMcSixBFctXQn7UA7vyFkH64zvT_OC_Pr29efVdb26-35ztVzVTkpVahBKdQvhXbOwreXIOYBVXHklpG1oxyQCk9zbzoNsuKNOe-CSa9RWgWPigtTTvz2szZDCBtLBRAjmerkyA-SCu2So4AuuRLM_8vOJdynmnLD7JzFqjn3N2Ne89B2Fj5Mw7OwG_Qt-CjoCn04XhKf-T0hobIiux43hWpvGLER7hJoJwrHFPmAy2QXcOvSj4IrxMfzvgL-VQZQB</recordid><startdate>20020222</startdate><enddate>20020222</enddate><creator>Dramsi, Shaynoor</creator><creator>Scheid, Michael P.</creator><creator>Maiti, Arpita</creator><creator>Hojabrpour, Payman</creator><creator>Chen, Xianming</creator><creator>Schubert, Kathryn</creator><creator>Goodlett, David R.</creator><creator>Aebersold, Ruedi</creator><creator>Duronio, Vincent</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>1XC</scope><scope>VOOES</scope></search><sort><creationdate>20020222</creationdate><title>Identification of a Novel Phosphorylation Site, Ser-170, as a Regulator of Bad Pro-apoptotic Activity</title><author>Dramsi, Shaynoor ; Scheid, Michael P. ; Maiti, Arpita ; Hojabrpour, Payman ; Chen, Xianming ; Schubert, Kathryn ; Goodlett, David R. ; Aebersold, Ruedi ; Duronio, Vincent</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c445t-a355f63dc86b9b2e22aab525d534b80f14ea142dbfda482c0c7da2427e7b5ac13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Amino Acid Substitution</topic><topic>Animals</topic><topic>Apoptosis</topic><topic>Apoptosis - physiology</topic><topic>bcl-Associated Death Protein</topic><topic>bcl-X Protein</topic><topic>Carrier Proteins</topic><topic>Carrier Proteins - chemistry</topic><topic>Carrier Proteins - metabolism</topic><topic>Chlorocebus aethiops</topic><topic>Cloning, Molecular</topic><topic>COS Cells</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Genes, bcl-2</topic><topic>Humans</topic><topic>Life Sciences</topic><topic>Mice</topic><topic>Mutagenesis, Site-Directed</topic><topic>Phosphorylation</topic><topic>Phosphoserine</topic><topic>Phosphoserine - metabolism</topic><topic>Poly(ADP-ribose) Polymerases</topic><topic>Poly(ADP-ribose) Polymerases - metabolism</topic><topic>Proto-Oncogene Proteins c-bcl-2</topic><topic>Proto-Oncogene Proteins c-bcl-2 - metabolism</topic><topic>Recombinant Proteins</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - metabolism</topic><topic>Serine</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dramsi, Shaynoor</creatorcontrib><creatorcontrib>Scheid, Michael P.</creatorcontrib><creatorcontrib>Maiti, Arpita</creatorcontrib><creatorcontrib>Hojabrpour, Payman</creatorcontrib><creatorcontrib>Chen, Xianming</creatorcontrib><creatorcontrib>Schubert, Kathryn</creatorcontrib><creatorcontrib>Goodlett, David R.</creatorcontrib><creatorcontrib>Aebersold, Ruedi</creatorcontrib><creatorcontrib>Duronio, Vincent</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>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dramsi, Shaynoor</au><au>Scheid, Michael P.</au><au>Maiti, Arpita</au><au>Hojabrpour, Payman</au><au>Chen, Xianming</au><au>Schubert, Kathryn</au><au>Goodlett, David R.</au><au>Aebersold, Ruedi</au><au>Duronio, Vincent</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of a Novel Phosphorylation Site, Ser-170, as a Regulator of Bad Pro-apoptotic Activity</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2002-02-22</date><risdate>2002</risdate><volume>277</volume><issue>8</issue><spage>6399</spage><epage>6405</epage><pages>6399-6405</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Bad is a pro-apoptotic member of the Bcl-2 family of proteins that is thought to exert a death-promoting effect by heterodimerization with Bcl-XL, nullifying its anti-apoptotic activity. Growth factors may promote cell survival at least partially through phosphorylation of Bad at one or more of Ser-112, -136, or -155. Our previous work showed that Bad is also phosphorylated in response to cytokines at another site, which we now identify as Ser-170. The functional role of this novel phosphorylation site was assessed by site-directed mutagenesis and analysis of the pro-apoptotic function of Bad in transiently transfected HEK293 and COS-7 cells or by stable expression in the cytokine-dependent cell line, MC/9. In general, mutation of Ser-170 to Ala results in a protein with increased ability to induce apoptosis, similar to the S112A mutant. Mutation of Ser-170 to Asp, mimicking a constitutively phosphorylated site, results in a protein that is virtually unable to induce apoptosis. Similarly, the S112A/S170D double mutant does not cause apoptosis in HEK293 and MC/9 cell lines. These data strongly suggest that phosphorylation of Bad at Ser-170 is a critical event in blocking the pro-apoptotic activity of Bad.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>11717309</pmid><doi>10.1074/jbc.M109990200</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 2002-02, Vol.277 (8), p.6399-6405 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_hal_primary_oai_HAL_pasteur_03262538v1 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Amino Acid Substitution Animals Apoptosis Apoptosis - physiology bcl-Associated Death Protein bcl-X Protein Carrier Proteins Carrier Proteins - chemistry Carrier Proteins - metabolism Chlorocebus aethiops Cloning, Molecular COS Cells Escherichia coli Escherichia coli - genetics Genes, bcl-2 Humans Life Sciences Mice Mutagenesis, Site-Directed Phosphorylation Phosphoserine Phosphoserine - metabolism Poly(ADP-ribose) Polymerases Poly(ADP-ribose) Polymerases - metabolism Proto-Oncogene Proteins c-bcl-2 Proto-Oncogene Proteins c-bcl-2 - metabolism Recombinant Proteins Recombinant Proteins - chemistry Recombinant Proteins - metabolism Serine Transfection |
| title | Identification of a Novel Phosphorylation Site, Ser-170, as a Regulator of Bad Pro-apoptotic Activity |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-13T13%3A09%3A44IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-hal_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Identification%20of%20a%20Novel%20Phosphorylation%20Site,%20Ser-170,%20as%20a%20Regulator%20of%20Bad%20Pro-apoptotic%20Activity&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Dramsi,%20Shaynoor&rft.date=2002-02-22&rft.volume=277&rft.issue=8&rft.spage=6399&rft.epage=6405&rft.pages=6399-6405&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.M109990200&rft_dat=%3Chal_cross%3Eoai_HAL_pasteur_03262538v1%3C/hal_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_id=info:pmid/11717309&rft_els_id=S0021925819824201&rfr_iscdi=true |