Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome
Holt-Oram syndrome is caused by mutations in TBX5, a member of the T-box gene family. In order to identify DNA sequences to which the TBX5 protein binds, we have performed an in vitro binding site selection assay. We have identified an 8 bp core sequence that is part of the Brachyury consensus-bindi...
Gespeichert in:
| Veröffentlicht in: | Human molecular genetics 2001-09, Vol.10 (18), p.1983-1994 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1994 |
|---|---|
| container_issue | 18 |
| container_start_page | 1983 |
| container_title | Human molecular genetics |
| container_volume | 10 |
| creator | GHOSH, Tushar K PACKHAM, Elizabeth A BONSER, Andrew J ROBINSON, Thelma E CROSS, Stephen J BROOK, J. David |
| description | Holt-Oram syndrome is caused by mutations in TBX5, a member of the T-box gene family. In order to identify DNA sequences to which the TBX5 protein binds, we have performed an in vitro binding site selection assay. We have identified an 8 bp core sequence that is part of the Brachyury consensus-binding site. We show that TBX5 binds to the full palindromic Brachyury binding site and to the half-palindrome, whereas Brachyury does not bind to the TBX5 site. Amino acids 1-237 of TBX5 are required for DNA binding. Analysis of the effects of specific substitution mutations that arise in Holt-Oram patients indicates that G80R and R237Q eliminate binding to the target site. DNA database analysis reveals that target sites are present in the upstream regions of several cardiac-expressed genes including cardiac alpha actin, atrial natriuretic factor, cardiac myosin heavy chain alpha, cardiac myosin heavy chain beta, myosin light chain 1A, myosin light chain 1V and Nkx2.5. Cell transfection studies demonstrate that TBX5 activates the transcription of an atrial natriuretic factor reporter construct and this effect is significantly reduced by deletion of the TBX5 binding site. |
| doi_str_mv | 10.1093/hmg/10.18.1983 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_71170314</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>71170314</sourcerecordid><originalsourceid>FETCH-LOGICAL-c392t-3e7e401ff7603f122b906e499a7870b59aa2eb64dd536e2f3df93fa4e2d635543</originalsourceid><addsrcrecordid>eNqF0DtPwzAQB3ALgaA8VkbkBba0dvyIPULFS0JiKRJbdEnO1CgPsJ2hfHpSqMTIcPINvzud_4ScczbnzIrFuntbbHsz59aIPTLjUrMsZ0bskxmzWmbaMn1EjmN8Z4xrKYpDcsS5UkoLNSOwXEOAOmHwX5D80NPB0bRGurp5VbTyfeP7Nxp9Qgp9MxW0m-jjVnVj-pmIk4dEaxgj0oehTdlzgI7GTd-EocNTcuCgjXi2e0_Iy93tavmQPT3fPy6vn7Ja2DxlAguUjDtXaCYcz_NquhultVCYglXKAuRYadk0SmjMnWicFQ4k5s30ESXFCbn63fsRhs8RYyo7H2tsW-hxGGNZcF4wwf-H3PDcSGUnOP-FdRhiDOjKj-A7CJuSs3Kbfjml_9Obcpv-NHCx2zxWHTZ_fBf3BC53AGINrQvQ1z7-OcmV1caIb-o1jTA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>18128459</pqid></control><display><type>article</type><title>Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome</title><source>MEDLINE</source><source>Oxford University Press Journals All Titles (1996-Current)</source><source>EZB-FREE-00999 freely available EZB journals</source><creator>GHOSH, Tushar K ; PACKHAM, Elizabeth A ; BONSER, Andrew J ; ROBINSON, Thelma E ; CROSS, Stephen J ; BROOK, J. David</creator><creatorcontrib>GHOSH, Tushar K ; PACKHAM, Elizabeth A ; BONSER, Andrew J ; ROBINSON, Thelma E ; CROSS, Stephen J ; BROOK, J. David</creatorcontrib><description>Holt-Oram syndrome is caused by mutations in TBX5, a member of the T-box gene family. In order to identify DNA sequences to which the TBX5 protein binds, we have performed an in vitro binding site selection assay. We have identified an 8 bp core sequence that is part of the Brachyury consensus-binding site. We show that TBX5 binds to the full palindromic Brachyury binding site and to the half-palindrome, whereas Brachyury does not bind to the TBX5 site. Amino acids 1-237 of TBX5 are required for DNA binding. Analysis of the effects of specific substitution mutations that arise in Holt-Oram patients indicates that G80R and R237Q eliminate binding to the target site. DNA database analysis reveals that target sites are present in the upstream regions of several cardiac-expressed genes including cardiac alpha actin, atrial natriuretic factor, cardiac myosin heavy chain alpha, cardiac myosin heavy chain beta, myosin light chain 1A, myosin light chain 1V and Nkx2.5. Cell transfection studies demonstrate that TBX5 activates the transcription of an atrial natriuretic factor reporter construct and this effect is significantly reduced by deletion of the TBX5 binding site.</description><identifier>ISSN: 0964-6906</identifier><identifier>ISSN: 1460-2083</identifier><identifier>EISSN: 1460-2083</identifier><identifier>DOI: 10.1093/hmg/10.18.1983</identifier><identifier>PMID: 11555635</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>Abnormalities, Multiple - genetics ; Abnormalities, Multiple - pathology ; Animals ; atrial natriuretic factor ; Atrial Natriuretic Factor - genetics ; Base Sequence ; Binding Sites - genetics ; Binding, Competitive ; Biological and medical sciences ; Brachyury gene ; Cell Line ; COS Cells ; Fetal Proteins ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation ; Genes, Reporter - genetics ; Heart Defects, Congenital - pathology ; Heart Ventricles - embryology ; Heart Ventricles - metabolism ; Holt-Oram syndrome ; In Situ Hybridization ; Limb Deformities, Congenital - pathology ; Mice ; Molecular and cellular biology ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; Myocardium - metabolism ; Oligonucleotides - genetics ; Oligonucleotides - metabolism ; Plasmids - genetics ; Promoter Regions, Genetic - genetics ; Sequence Homology, Nucleic Acid ; Syndrome ; T-Box Domain Proteins - genetics ; T-Box Domain Proteins - metabolism ; TBX5 gene ; TBX5 protein</subject><ispartof>Human molecular genetics, 2001-09, Vol.10 (18), p.1983-1994</ispartof><rights>2002 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c392t-3e7e401ff7603f122b906e499a7870b59aa2eb64dd536e2f3df93fa4e2d635543</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=14159688$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11555635$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>GHOSH, Tushar K</creatorcontrib><creatorcontrib>PACKHAM, Elizabeth A</creatorcontrib><creatorcontrib>BONSER, Andrew J</creatorcontrib><creatorcontrib>ROBINSON, Thelma E</creatorcontrib><creatorcontrib>CROSS, Stephen J</creatorcontrib><creatorcontrib>BROOK, J. David</creatorcontrib><title>Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome</title><title>Human molecular genetics</title><addtitle>Hum Mol Genet</addtitle><description>Holt-Oram syndrome is caused by mutations in TBX5, a member of the T-box gene family. In order to identify DNA sequences to which the TBX5 protein binds, we have performed an in vitro binding site selection assay. We have identified an 8 bp core sequence that is part of the Brachyury consensus-binding site. We show that TBX5 binds to the full palindromic Brachyury binding site and to the half-palindrome, whereas Brachyury does not bind to the TBX5 site. Amino acids 1-237 of TBX5 are required for DNA binding. Analysis of the effects of specific substitution mutations that arise in Holt-Oram patients indicates that G80R and R237Q eliminate binding to the target site. DNA database analysis reveals that target sites are present in the upstream regions of several cardiac-expressed genes including cardiac alpha actin, atrial natriuretic factor, cardiac myosin heavy chain alpha, cardiac myosin heavy chain beta, myosin light chain 1A, myosin light chain 1V and Nkx2.5. Cell transfection studies demonstrate that TBX5 activates the transcription of an atrial natriuretic factor reporter construct and this effect is significantly reduced by deletion of the TBX5 binding site.</description><subject>Abnormalities, Multiple - genetics</subject><subject>Abnormalities, Multiple - pathology</subject><subject>Animals</subject><subject>atrial natriuretic factor</subject><subject>Atrial Natriuretic Factor - genetics</subject><subject>Base Sequence</subject><subject>Binding Sites - genetics</subject><subject>Binding, Competitive</subject><subject>Biological and medical sciences</subject><subject>Brachyury gene</subject><subject>Cell Line</subject><subject>COS Cells</subject><subject>Fetal Proteins</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation</subject><subject>Genes, Reporter - genetics</subject><subject>Heart Defects, Congenital - pathology</subject><subject>Heart Ventricles - embryology</subject><subject>Heart Ventricles - metabolism</subject><subject>Holt-Oram syndrome</subject><subject>In Situ Hybridization</subject><subject>Limb Deformities, Congenital - pathology</subject><subject>Mice</subject><subject>Molecular and cellular biology</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>Mutation, Missense</subject><subject>Myocardium - metabolism</subject><subject>Oligonucleotides - genetics</subject><subject>Oligonucleotides - metabolism</subject><subject>Plasmids - genetics</subject><subject>Promoter Regions, Genetic - genetics</subject><subject>Sequence Homology, Nucleic Acid</subject><subject>Syndrome</subject><subject>T-Box Domain Proteins - genetics</subject><subject>T-Box Domain Proteins - metabolism</subject><subject>TBX5 gene</subject><subject>TBX5 protein</subject><issn>0964-6906</issn><issn>1460-2083</issn><issn>1460-2083</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0DtPwzAQB3ALgaA8VkbkBba0dvyIPULFS0JiKRJbdEnO1CgPsJ2hfHpSqMTIcPINvzud_4ScczbnzIrFuntbbHsz59aIPTLjUrMsZ0bskxmzWmbaMn1EjmN8Z4xrKYpDcsS5UkoLNSOwXEOAOmHwX5D80NPB0bRGurp5VbTyfeP7Nxp9Qgp9MxW0m-jjVnVj-pmIk4dEaxgj0oehTdlzgI7GTd-EocNTcuCgjXi2e0_Iy93tavmQPT3fPy6vn7Ja2DxlAguUjDtXaCYcz_NquhultVCYglXKAuRYadk0SmjMnWicFQ4k5s30ESXFCbn63fsRhs8RYyo7H2tsW-hxGGNZcF4wwf-H3PDcSGUnOP-FdRhiDOjKj-A7CJuSs3Kbfjml_9Obcpv-NHCx2zxWHTZ_fBf3BC53AGINrQvQ1z7-OcmV1caIb-o1jTA</recordid><startdate>20010901</startdate><enddate>20010901</enddate><creator>GHOSH, Tushar K</creator><creator>PACKHAM, Elizabeth A</creator><creator>BONSER, Andrew J</creator><creator>ROBINSON, Thelma E</creator><creator>CROSS, Stephen J</creator><creator>BROOK, J. David</creator><general>Oxford University Press</general><scope>IQODW</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>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20010901</creationdate><title>Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome</title><author>GHOSH, Tushar K ; PACKHAM, Elizabeth A ; BONSER, Andrew J ; ROBINSON, Thelma E ; CROSS, Stephen J ; BROOK, J. David</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c392t-3e7e401ff7603f122b906e499a7870b59aa2eb64dd536e2f3df93fa4e2d635543</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Abnormalities, Multiple - genetics</topic><topic>Abnormalities, Multiple - pathology</topic><topic>Animals</topic><topic>atrial natriuretic factor</topic><topic>Atrial Natriuretic Factor - genetics</topic><topic>Base Sequence</topic><topic>Binding Sites - genetics</topic><topic>Binding, Competitive</topic><topic>Biological and medical sciences</topic><topic>Brachyury gene</topic><topic>Cell Line</topic><topic>COS Cells</topic><topic>Fetal Proteins</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation</topic><topic>Genes, Reporter - genetics</topic><topic>Heart Defects, Congenital - pathology</topic><topic>Heart Ventricles - embryology</topic><topic>Heart Ventricles - metabolism</topic><topic>Holt-Oram syndrome</topic><topic>In Situ Hybridization</topic><topic>Limb Deformities, Congenital - pathology</topic><topic>Mice</topic><topic>Molecular and cellular biology</topic><topic>Molecular Sequence Data</topic><topic>Mutation</topic><topic>Mutation, Missense</topic><topic>Myocardium - metabolism</topic><topic>Oligonucleotides - genetics</topic><topic>Oligonucleotides - metabolism</topic><topic>Plasmids - genetics</topic><topic>Promoter Regions, Genetic - genetics</topic><topic>Sequence Homology, Nucleic Acid</topic><topic>Syndrome</topic><topic>T-Box Domain Proteins - genetics</topic><topic>T-Box Domain Proteins - metabolism</topic><topic>TBX5 gene</topic><topic>TBX5 protein</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>GHOSH, Tushar K</creatorcontrib><creatorcontrib>PACKHAM, Elizabeth A</creatorcontrib><creatorcontrib>BONSER, Andrew J</creatorcontrib><creatorcontrib>ROBINSON, Thelma E</creatorcontrib><creatorcontrib>CROSS, Stephen J</creatorcontrib><creatorcontrib>BROOK, J. David</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Human molecular genetics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>GHOSH, Tushar K</au><au>PACKHAM, Elizabeth A</au><au>BONSER, Andrew J</au><au>ROBINSON, Thelma E</au><au>CROSS, Stephen J</au><au>BROOK, J. David</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome</atitle><jtitle>Human molecular genetics</jtitle><addtitle>Hum Mol Genet</addtitle><date>2001-09-01</date><risdate>2001</risdate><volume>10</volume><issue>18</issue><spage>1983</spage><epage>1994</epage><pages>1983-1994</pages><issn>0964-6906</issn><issn>1460-2083</issn><eissn>1460-2083</eissn><abstract>Holt-Oram syndrome is caused by mutations in TBX5, a member of the T-box gene family. In order to identify DNA sequences to which the TBX5 protein binds, we have performed an in vitro binding site selection assay. We have identified an 8 bp core sequence that is part of the Brachyury consensus-binding site. We show that TBX5 binds to the full palindromic Brachyury binding site and to the half-palindrome, whereas Brachyury does not bind to the TBX5 site. Amino acids 1-237 of TBX5 are required for DNA binding. Analysis of the effects of specific substitution mutations that arise in Holt-Oram patients indicates that G80R and R237Q eliminate binding to the target site. DNA database analysis reveals that target sites are present in the upstream regions of several cardiac-expressed genes including cardiac alpha actin, atrial natriuretic factor, cardiac myosin heavy chain alpha, cardiac myosin heavy chain beta, myosin light chain 1A, myosin light chain 1V and Nkx2.5. Cell transfection studies demonstrate that TBX5 activates the transcription of an atrial natriuretic factor reporter construct and this effect is significantly reduced by deletion of the TBX5 binding site.</abstract><cop>Oxford</cop><pub>Oxford University Press</pub><pmid>11555635</pmid><doi>10.1093/hmg/10.18.1983</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0964-6906 |
| ispartof | Human molecular genetics, 2001-09, Vol.10 (18), p.1983-1994 |
| issn | 0964-6906 1460-2083 1460-2083 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_71170314 |
| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals |
| subjects | Abnormalities, Multiple - genetics Abnormalities, Multiple - pathology Animals atrial natriuretic factor Atrial Natriuretic Factor - genetics Base Sequence Binding Sites - genetics Binding, Competitive Biological and medical sciences Brachyury gene Cell Line COS Cells Fetal Proteins Fundamental and applied biological sciences. Psychology Gene Expression Regulation Genes, Reporter - genetics Heart Defects, Congenital - pathology Heart Ventricles - embryology Heart Ventricles - metabolism Holt-Oram syndrome In Situ Hybridization Limb Deformities, Congenital - pathology Mice Molecular and cellular biology Molecular Sequence Data Mutation Mutation, Missense Myocardium - metabolism Oligonucleotides - genetics Oligonucleotides - metabolism Plasmids - genetics Promoter Regions, Genetic - genetics Sequence Homology, Nucleic Acid Syndrome T-Box Domain Proteins - genetics T-Box Domain Proteins - metabolism TBX5 gene TBX5 protein |
| title | Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-06T08%3A52%3A26IST&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=Characterization%20of%20the%20TBX5%20binding%20site%20and%20analysis%20of%20mutations%20that%20cause%20Holt-Oram%20syndrome&rft.jtitle=Human%20molecular%20genetics&rft.au=GHOSH,%20Tushar%20K&rft.date=2001-09-01&rft.volume=10&rft.issue=18&rft.spage=1983&rft.epage=1994&rft.pages=1983-1994&rft.issn=0964-6906&rft.eissn=1460-2083&rft_id=info:doi/10.1093/hmg/10.18.1983&rft_dat=%3Cproquest_cross%3E71170314%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=18128459&rft_id=info:pmid/11555635&rfr_iscdi=true |