Observation of unphosphorylated STAT3 core protein binding to target dsDNA by PEMSA and X-ray crystallography
► Stoichiometric uSTAT3 interaction with M67 DNA. ► Interaction is weaker compared to pSTAT3 M67 DNA interaction. ► Report of a novel protein electrophoretic mobility shift assay (PEMSA). The STAT3 transcription factor plays a central role in a wide range of cancer types where it is over-expressed....
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| Veröffentlicht in: | FEBS letters 2013-04, Vol.587 (7), p.833-839 |
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| creator | Nkansah, Edwin Shah, Rahi Collie, Gavin W. Parkinson, Gary N. Palmer, Jonathan Rahman, Khondaker M. Bui, Tam T. Drake, Alex F. Husby, Jarmila Neidle, Stephen Zinzalla, Giovanna Thurston, David E. Wilderspin, Andrew F. |
| description | ► Stoichiometric uSTAT3 interaction with M67 DNA. ► Interaction is weaker compared to pSTAT3 M67 DNA interaction. ► Report of a novel protein electrophoretic mobility shift assay (PEMSA).
The STAT3 transcription factor plays a central role in a wide range of cancer types where it is over-expressed. Previously, phosphorylation of this protein was thought to be a prerequisite for direct binding to DNA. However, we have now shown complete binding of a purified unphosphorylated STAT3 (uSTAT3) core directly to M67 DNA, the high affinity STAT3 target DNA sequence, by a protein electrophoretic mobility shift assay (PEMSA). Binding to M67 DNA was inhibited by addition of increasing concentrations of a phosphotyrosyl peptide. X-ray crystallography demonstrates one mode of binding that is similar to that known for the STAT3 core phosphorylated at Y705.
pSTAT3βtcandpSTAT3βtcbindbymolecular sieving(View interaction) |
| doi_str_mv | 10.1016/j.febslet.2013.01.065 |
| format | Article |
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The STAT3 transcription factor plays a central role in a wide range of cancer types where it is over-expressed. Previously, phosphorylation of this protein was thought to be a prerequisite for direct binding to DNA. However, we have now shown complete binding of a purified unphosphorylated STAT3 (uSTAT3) core directly to M67 DNA, the high affinity STAT3 target DNA sequence, by a protein electrophoretic mobility shift assay (PEMSA). Binding to M67 DNA was inhibited by addition of increasing concentrations of a phosphotyrosyl peptide. X-ray crystallography demonstrates one mode of binding that is similar to that known for the STAT3 core phosphorylated at Y705.
pSTAT3βtcandpSTAT3βtcbindbymolecular sieving(View interaction)</description><identifier>ISSN: 0014-5793</identifier><identifier>ISSN: 1873-3468</identifier><identifier>EISSN: 1873-3468</identifier><identifier>DOI: 10.1016/j.febslet.2013.01.065</identifier><identifier>PMID: 23434585</identifier><language>eng</language><publisher>England: Elsevier B.V</publisher><subject>Animals ; Binding Sites ; Binding, Competitive ; Circular Dichroism ; Crystallography, X-Ray ; DNA - chemistry ; DNA - genetics ; DNA - metabolism ; Electrophoretic Mobility Shift Assay - methods ; Humans ; Luminescent Proteins - genetics ; Luminescent Proteins - metabolism ; M67 ; Models, Molecular ; Nucleic Acid Conformation ; PEMSA ; Phosphopeptides - chemistry ; Phosphopeptides - metabolism ; Phosphorylation ; Protein Binding ; Protein Structure, Secondary ; Protein Structure, Tertiary ; SH2 ; Signal transducer and activator of transcription 3 ; src homology 2 domain ; STAT3 ; STAT3 Transcription Factor - chemistry ; STAT3 Transcription Factor - genetics ; STAT3 Transcription Factor - metabolism ; STAT3βtc ; the STAT3 core consisting of amino acid residues 127–722 ; Tyrosine - chemistry ; Tyrosine - genetics ; Tyrosine - metabolism ; X-ray crystallography ; yellow variant of eGFP, the enhanced green fluorescent protein ; YFP</subject><ispartof>FEBS letters, 2013-04, Vol.587 (7), p.833-839</ispartof><rights>2013 Federation of European Biochemical Societies</rights><rights>FEBS Letters 587 (2013) 1873-3468 © 2015 Federation of European Biochemical Societies</rights><rights>Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1016%2Fj.febslet.2013.01.065$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0014579313001105$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,776,780,1411,1427,3537,27901,27902,45550,45551,46384,46808,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23434585$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nkansah, Edwin</creatorcontrib><creatorcontrib>Shah, Rahi</creatorcontrib><creatorcontrib>Collie, Gavin W.</creatorcontrib><creatorcontrib>Parkinson, Gary N.</creatorcontrib><creatorcontrib>Palmer, Jonathan</creatorcontrib><creatorcontrib>Rahman, Khondaker M.</creatorcontrib><creatorcontrib>Bui, Tam T.</creatorcontrib><creatorcontrib>Drake, Alex F.</creatorcontrib><creatorcontrib>Husby, Jarmila</creatorcontrib><creatorcontrib>Neidle, Stephen</creatorcontrib><creatorcontrib>Zinzalla, Giovanna</creatorcontrib><creatorcontrib>Thurston, David E.</creatorcontrib><creatorcontrib>Wilderspin, Andrew F.</creatorcontrib><title>Observation of unphosphorylated STAT3 core protein binding to target dsDNA by PEMSA and X-ray crystallography</title><title>FEBS letters</title><addtitle>FEBS Lett</addtitle><description>► Stoichiometric uSTAT3 interaction with M67 DNA. ► Interaction is weaker compared to pSTAT3 M67 DNA interaction. ► Report of a novel protein electrophoretic mobility shift assay (PEMSA).
The STAT3 transcription factor plays a central role in a wide range of cancer types where it is over-expressed. Previously, phosphorylation of this protein was thought to be a prerequisite for direct binding to DNA. However, we have now shown complete binding of a purified unphosphorylated STAT3 (uSTAT3) core directly to M67 DNA, the high affinity STAT3 target DNA sequence, by a protein electrophoretic mobility shift assay (PEMSA). Binding to M67 DNA was inhibited by addition of increasing concentrations of a phosphotyrosyl peptide. X-ray crystallography demonstrates one mode of binding that is similar to that known for the STAT3 core phosphorylated at Y705.
pSTAT3βtcandpSTAT3βtcbindbymolecular sieving(View interaction)</description><subject>Animals</subject><subject>Binding Sites</subject><subject>Binding, Competitive</subject><subject>Circular Dichroism</subject><subject>Crystallography, X-Ray</subject><subject>DNA - chemistry</subject><subject>DNA - genetics</subject><subject>DNA - metabolism</subject><subject>Electrophoretic Mobility Shift Assay - methods</subject><subject>Humans</subject><subject>Luminescent Proteins - genetics</subject><subject>Luminescent Proteins - metabolism</subject><subject>M67</subject><subject>Models, Molecular</subject><subject>Nucleic Acid Conformation</subject><subject>PEMSA</subject><subject>Phosphopeptides - chemistry</subject><subject>Phosphopeptides - metabolism</subject><subject>Phosphorylation</subject><subject>Protein Binding</subject><subject>Protein Structure, Secondary</subject><subject>Protein Structure, Tertiary</subject><subject>SH2</subject><subject>Signal transducer and activator of transcription 3</subject><subject>src homology 2 domain</subject><subject>STAT3</subject><subject>STAT3 Transcription Factor - chemistry</subject><subject>STAT3 Transcription Factor - genetics</subject><subject>STAT3 Transcription Factor - metabolism</subject><subject>STAT3βtc</subject><subject>the STAT3 core consisting of amino acid residues 127–722</subject><subject>Tyrosine - chemistry</subject><subject>Tyrosine - genetics</subject><subject>Tyrosine - metabolism</subject><subject>X-ray crystallography</subject><subject>yellow variant of eGFP, the enhanced green fluorescent protein</subject><subject>YFP</subject><issn>0014-5793</issn><issn>1873-3468</issn><issn>1873-3468</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkcFuEzEQhi0EoqHwCCAfuWxqe-yN94RCSVukQpESJG6W155NHW12g-0U7duzqwSOiMNoxvKnX6P5CHnL2ZwzXl7t5g3WqcU8F4zDnPE5K9UzMuN6AQXIUj8nM8a4LNSiggvyKqUdG9-aVy_JhQAJUmk1I_uHOmF8sjn0He0beuwOj30aKw6tzejperPcAHV9RHqIfcbQ0Tp0PnRbmnuabdxipj59-rqk9UC_rb6sl9R2nv4ooh2oi0PKtm37bbSHx-E1edHYNuGbc78k329Wm-u74v7h9vP18r5ACVwVQjsEpdHbsUPNUJTSey-drRyUylqnrQBhFwvVNEJ7XzPnGSuhUtgoALgk70-548o_j5iy2YfksG1th_0xGQ5SSw1Ci_9AeVUKreWEvjujx3qP3hxi2Ns4mD_XHIG7E_ArtDj8_efMTMrMzpyVmUmZYdyMyszN6qNYT6YmURzGibMp6sMpCsczPQWMJrmAnUMfIrpsfB_-nQu_AVIRpFs</recordid><startdate>20130402</startdate><enddate>20130402</enddate><creator>Nkansah, Edwin</creator><creator>Shah, Rahi</creator><creator>Collie, Gavin W.</creator><creator>Parkinson, Gary N.</creator><creator>Palmer, Jonathan</creator><creator>Rahman, Khondaker M.</creator><creator>Bui, Tam T.</creator><creator>Drake, Alex F.</creator><creator>Husby, Jarmila</creator><creator>Neidle, Stephen</creator><creator>Zinzalla, Giovanna</creator><creator>Thurston, David E.</creator><creator>Wilderspin, Andrew F.</creator><general>Elsevier B.V</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>7X8</scope><scope>7TM</scope></search><sort><creationdate>20130402</creationdate><title>Observation of unphosphorylated STAT3 core protein binding to target dsDNA by PEMSA and X-ray crystallography</title><author>Nkansah, Edwin ; Shah, Rahi ; Collie, Gavin W. ; Parkinson, Gary N. ; Palmer, Jonathan ; Rahman, Khondaker M. ; Bui, Tam T. ; Drake, Alex F. ; Husby, Jarmila ; Neidle, Stephen ; Zinzalla, Giovanna ; Thurston, David E. ; Wilderspin, Andrew F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-e4315-28ce358edace33b0e264ddd4ca9c365aac8a232a775ff28ddb0cd006395ef5333</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>Binding Sites</topic><topic>Binding, Competitive</topic><topic>Circular Dichroism</topic><topic>Crystallography, X-Ray</topic><topic>DNA - chemistry</topic><topic>DNA - genetics</topic><topic>DNA - metabolism</topic><topic>Electrophoretic Mobility Shift Assay - methods</topic><topic>Humans</topic><topic>Luminescent Proteins - genetics</topic><topic>Luminescent Proteins - metabolism</topic><topic>M67</topic><topic>Models, Molecular</topic><topic>Nucleic Acid Conformation</topic><topic>PEMSA</topic><topic>Phosphopeptides - chemistry</topic><topic>Phosphopeptides - metabolism</topic><topic>Phosphorylation</topic><topic>Protein Binding</topic><topic>Protein Structure, Secondary</topic><topic>Protein Structure, Tertiary</topic><topic>SH2</topic><topic>Signal transducer and activator of transcription 3</topic><topic>src homology 2 domain</topic><topic>STAT3</topic><topic>STAT3 Transcription Factor - chemistry</topic><topic>STAT3 Transcription Factor - genetics</topic><topic>STAT3 Transcription Factor - metabolism</topic><topic>STAT3βtc</topic><topic>the STAT3 core consisting of amino acid residues 127–722</topic><topic>Tyrosine - chemistry</topic><topic>Tyrosine - genetics</topic><topic>Tyrosine - metabolism</topic><topic>X-ray crystallography</topic><topic>yellow variant of eGFP, the enhanced green fluorescent protein</topic><topic>YFP</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nkansah, Edwin</creatorcontrib><creatorcontrib>Shah, Rahi</creatorcontrib><creatorcontrib>Collie, Gavin W.</creatorcontrib><creatorcontrib>Parkinson, Gary N.</creatorcontrib><creatorcontrib>Palmer, Jonathan</creatorcontrib><creatorcontrib>Rahman, Khondaker M.</creatorcontrib><creatorcontrib>Bui, Tam T.</creatorcontrib><creatorcontrib>Drake, Alex F.</creatorcontrib><creatorcontrib>Husby, Jarmila</creatorcontrib><creatorcontrib>Neidle, Stephen</creatorcontrib><creatorcontrib>Zinzalla, Giovanna</creatorcontrib><creatorcontrib>Thurston, David E.</creatorcontrib><creatorcontrib>Wilderspin, Andrew F.</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>MEDLINE - Academic</collection><collection>Nucleic Acids Abstracts</collection><jtitle>FEBS letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nkansah, Edwin</au><au>Shah, Rahi</au><au>Collie, Gavin W.</au><au>Parkinson, Gary N.</au><au>Palmer, Jonathan</au><au>Rahman, Khondaker M.</au><au>Bui, Tam T.</au><au>Drake, Alex F.</au><au>Husby, Jarmila</au><au>Neidle, Stephen</au><au>Zinzalla, Giovanna</au><au>Thurston, David E.</au><au>Wilderspin, Andrew F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Observation of unphosphorylated STAT3 core protein binding to target dsDNA by PEMSA and X-ray crystallography</atitle><jtitle>FEBS letters</jtitle><addtitle>FEBS Lett</addtitle><date>2013-04-02</date><risdate>2013</risdate><volume>587</volume><issue>7</issue><spage>833</spage><epage>839</epage><pages>833-839</pages><issn>0014-5793</issn><issn>1873-3468</issn><eissn>1873-3468</eissn><abstract>► Stoichiometric uSTAT3 interaction with M67 DNA. ► Interaction is weaker compared to pSTAT3 M67 DNA interaction. ► Report of a novel protein electrophoretic mobility shift assay (PEMSA).
The STAT3 transcription factor plays a central role in a wide range of cancer types where it is over-expressed. Previously, phosphorylation of this protein was thought to be a prerequisite for direct binding to DNA. However, we have now shown complete binding of a purified unphosphorylated STAT3 (uSTAT3) core directly to M67 DNA, the high affinity STAT3 target DNA sequence, by a protein electrophoretic mobility shift assay (PEMSA). Binding to M67 DNA was inhibited by addition of increasing concentrations of a phosphotyrosyl peptide. X-ray crystallography demonstrates one mode of binding that is similar to that known for the STAT3 core phosphorylated at Y705.
pSTAT3βtcandpSTAT3βtcbindbymolecular sieving(View interaction)</abstract><cop>England</cop><pub>Elsevier B.V</pub><pmid>23434585</pmid><doi>10.1016/j.febslet.2013.01.065</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Binding Sites Binding, Competitive Circular Dichroism Crystallography, X-Ray DNA - chemistry DNA - genetics DNA - metabolism Electrophoretic Mobility Shift Assay - methods Humans Luminescent Proteins - genetics Luminescent Proteins - metabolism M67 Models, Molecular Nucleic Acid Conformation PEMSA Phosphopeptides - chemistry Phosphopeptides - metabolism Phosphorylation Protein Binding Protein Structure, Secondary Protein Structure, Tertiary SH2 Signal transducer and activator of transcription 3 src homology 2 domain STAT3 STAT3 Transcription Factor - chemistry STAT3 Transcription Factor - genetics STAT3 Transcription Factor - metabolism STAT3βtc the STAT3 core consisting of amino acid residues 127–722 Tyrosine - chemistry Tyrosine - genetics Tyrosine - metabolism X-ray crystallography yellow variant of eGFP, the enhanced green fluorescent protein YFP |
| title | Observation of unphosphorylated STAT3 core protein binding to target dsDNA by PEMSA and X-ray crystallography |
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