Cloning and Identification of Regulatory Sequences of the Human Thrombin Receptor Gene
Thrombin, via activation of vascular endothelial and smooth muscle cell thrombin receptors, modulates vascular wall healing. To understand the mechanisms that regulate human thrombin receptor (HTR) expression, we cloned and characterized the HTR gene. The HTR gene consists of Exon I, which contains...
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| Veröffentlicht in: | The Journal of biological chemistry 1996-10, Vol.271 (42), p.26320-26328 |
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| container_title | The Journal of biological chemistry |
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| creator | Li, F Baykal, D Horaist, C Yan, C N Carr, B N Rao, G N Runge, M S |
| description | Thrombin, via activation of vascular endothelial and smooth muscle cell thrombin receptors, modulates vascular wall healing.
To understand the mechanisms that regulate human thrombin receptor (HTR) expression, we cloned and characterized the HTR gene.
The HTR gene consists of Exon I, which contains the 5â²-regulatory region and 85 nucleotides of coding sequence; a ~15-kb intron;
and Exon II, which contains the remainder of the coding sequence and the entire 3â²-untranslated region. Multiple transcription
initiation sites were identified by S1 mapping and ribonuclease protection assay. DNA sequence analysis indicated the presence
of two SP-1-AP-2 consensus binding sequences, near or within the transcription initiation sites, and consensus binding sequences
for numerous regulatory proteins that potentially modulate HTR expression. Functional analysis of the HTR promoter was performed
by transfecting human microvascular endothelial cells with HTR promoter region-luciferase constructs. The highest level of
expression was obtained with a 0.7-kb promoter sequence and was progressively less with fragments of 0.54, 1.16, 1.6, and
~3.2 kb. The data presented in this report provide a foundation for further characterization of the HTR gene and the mechanisms
that regulate its expression within the blood vessel wall. |
| doi_str_mv | 10.1074/jbc.271.42.26320 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_15755815</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>15755815</sourcerecordid><originalsourceid>FETCH-LOGICAL-c3770-f8f116e7ac0485efa319e4aba9ad338c36b69a260eb6e8c6ffcd6b0eb6c31fb53</originalsourceid><addsrcrecordid>eNpVkM1Lw0AQxRdRaq3evQg5iLfU_cjH5ihF24IgaBVvy-5mttmS7NZsgvS_N7VFcC7DzHvzGH4IXRM8JThP7jdKT2lOpgmd0oxRfILGBHMWs5R8nqIxxpTEBU35OboIYYOHSgoyQiPOaUJ5OkYfs9o769aRdGW0LMF11lgtO-td5E30Cuu-lp1vd9EbfPXgNIT9vqsgWvSNdNGqan2jrBusGraDM5qDg0t0ZmQd4OrYJ-j96XE1W8TPL_Pl7OE51izPcWy4ISSDXGqc8BSMZKSARCpZyJIxrlmmskLSDIPKgOvMGF1maj9pRoxK2QTdHXK3rR_eC51obNBQ19KB74MgaZ6mnOyN-GDUrQ-hBSO2rW1kuxMEiz1KMaAUA0qRUPGLcji5OWb3qoHy7-DIbtBvD3pl19W3bUEo63UFzf-YH_kXfE0</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>15755815</pqid></control><display><type>article</type><title>Cloning and Identification of Regulatory Sequences of the Human Thrombin Receptor Gene</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>Li, F ; Baykal, D ; Horaist, C ; Yan, C N ; Carr, B N ; Rao, G N ; Runge, M S</creator><creatorcontrib>Li, F ; Baykal, D ; Horaist, C ; Yan, C N ; Carr, B N ; Rao, G N ; Runge, M S</creatorcontrib><description>Thrombin, via activation of vascular endothelial and smooth muscle cell thrombin receptors, modulates vascular wall healing.
To understand the mechanisms that regulate human thrombin receptor (HTR) expression, we cloned and characterized the HTR gene.
The HTR gene consists of Exon I, which contains the 5â²-regulatory region and 85 nucleotides of coding sequence; a ~15-kb intron;
and Exon II, which contains the remainder of the coding sequence and the entire 3â²-untranslated region. Multiple transcription
initiation sites were identified by S1 mapping and ribonuclease protection assay. DNA sequence analysis indicated the presence
of two SP-1-AP-2 consensus binding sequences, near or within the transcription initiation sites, and consensus binding sequences
for numerous regulatory proteins that potentially modulate HTR expression. Functional analysis of the HTR promoter was performed
by transfecting human microvascular endothelial cells with HTR promoter region-luciferase constructs. The highest level of
expression was obtained with a 0.7-kb promoter sequence and was progressively less with fragments of 0.54, 1.16, 1.6, and
~3.2 kb. The data presented in this report provide a foundation for further characterization of the HTR gene and the mechanisms
that regulate its expression within the blood vessel wall.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.271.42.26320</identifier><identifier>PMID: 8824285</identifier><language>eng</language><publisher>United States: American Society for Biochemistry and Molecular Biology</publisher><subject>Amino Acid Sequence ; Base Sequence ; Blotting, Southern ; Chromosome Mapping ; Cloning, Molecular ; DNA ; Exons ; Gene Deletion ; Humans ; Introns ; Microscopy, Electron ; Molecular Sequence Data ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Receptors, Thrombin - genetics ; Restriction Mapping ; Single-Strand Specific DNA and RNA Endonucleases - metabolism</subject><ispartof>The Journal of biological chemistry, 1996-10, Vol.271 (42), p.26320-26328</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3770-f8f116e7ac0485efa319e4aba9ad338c36b69a260eb6e8c6ffcd6b0eb6c31fb53</citedby><cites>FETCH-LOGICAL-c3770-f8f116e7ac0485efa319e4aba9ad338c36b69a260eb6e8c6ffcd6b0eb6c31fb53</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/8824285$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, F</creatorcontrib><creatorcontrib>Baykal, D</creatorcontrib><creatorcontrib>Horaist, C</creatorcontrib><creatorcontrib>Yan, C N</creatorcontrib><creatorcontrib>Carr, B N</creatorcontrib><creatorcontrib>Rao, G N</creatorcontrib><creatorcontrib>Runge, M S</creatorcontrib><title>Cloning and Identification of Regulatory Sequences of the Human Thrombin Receptor Gene</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Thrombin, via activation of vascular endothelial and smooth muscle cell thrombin receptors, modulates vascular wall healing.
To understand the mechanisms that regulate human thrombin receptor (HTR) expression, we cloned and characterized the HTR gene.
The HTR gene consists of Exon I, which contains the 5â²-regulatory region and 85 nucleotides of coding sequence; a ~15-kb intron;
and Exon II, which contains the remainder of the coding sequence and the entire 3â²-untranslated region. Multiple transcription
initiation sites were identified by S1 mapping and ribonuclease protection assay. DNA sequence analysis indicated the presence
of two SP-1-AP-2 consensus binding sequences, near or within the transcription initiation sites, and consensus binding sequences
for numerous regulatory proteins that potentially modulate HTR expression. Functional analysis of the HTR promoter was performed
by transfecting human microvascular endothelial cells with HTR promoter region-luciferase constructs. The highest level of
expression was obtained with a 0.7-kb promoter sequence and was progressively less with fragments of 0.54, 1.16, 1.6, and
~3.2 kb. The data presented in this report provide a foundation for further characterization of the HTR gene and the mechanisms
that regulate its expression within the blood vessel wall.</description><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>Blotting, Southern</subject><subject>Chromosome Mapping</subject><subject>Cloning, Molecular</subject><subject>DNA</subject><subject>Exons</subject><subject>Gene Deletion</subject><subject>Humans</subject><subject>Introns</subject><subject>Microscopy, Electron</subject><subject>Molecular Sequence Data</subject><subject>Polymerase Chain Reaction</subject><subject>Promoter Regions, Genetic</subject><subject>Receptors, Thrombin - genetics</subject><subject>Restriction Mapping</subject><subject>Single-Strand Specific DNA and RNA Endonucleases - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkM1Lw0AQxRdRaq3evQg5iLfU_cjH5ihF24IgaBVvy-5mttmS7NZsgvS_N7VFcC7DzHvzGH4IXRM8JThP7jdKT2lOpgmd0oxRfILGBHMWs5R8nqIxxpTEBU35OboIYYOHSgoyQiPOaUJ5OkYfs9o769aRdGW0LMF11lgtO-td5E30Cuu-lp1vd9EbfPXgNIT9vqsgWvSNdNGqan2jrBusGraDM5qDg0t0ZmQd4OrYJ-j96XE1W8TPL_Pl7OE51izPcWy4ISSDXGqc8BSMZKSARCpZyJIxrlmmskLSDIPKgOvMGF1maj9pRoxK2QTdHXK3rR_eC51obNBQ19KB74MgaZ6mnOyN-GDUrQ-hBSO2rW1kuxMEiz1KMaAUA0qRUPGLcji5OWb3qoHy7-DIbtBvD3pl19W3bUEo63UFzf-YH_kXfE0</recordid><startdate>19961018</startdate><enddate>19961018</enddate><creator>Li, F</creator><creator>Baykal, D</creator><creator>Horaist, C</creator><creator>Yan, C N</creator><creator>Carr, B N</creator><creator>Rao, G N</creator><creator>Runge, M S</creator><general>American Society for Biochemistry and Molecular Biology</general><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>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>19961018</creationdate><title>Cloning and Identification of Regulatory Sequences of the Human Thrombin Receptor Gene</title><author>Li, F ; Baykal, D ; Horaist, C ; Yan, C N ; Carr, B N ; Rao, G N ; Runge, M S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3770-f8f116e7ac0485efa319e4aba9ad338c36b69a260eb6e8c6ffcd6b0eb6c31fb53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Amino Acid Sequence</topic><topic>Base Sequence</topic><topic>Blotting, Southern</topic><topic>Chromosome Mapping</topic><topic>Cloning, Molecular</topic><topic>DNA</topic><topic>Exons</topic><topic>Gene Deletion</topic><topic>Humans</topic><topic>Introns</topic><topic>Microscopy, Electron</topic><topic>Molecular Sequence Data</topic><topic>Polymerase Chain Reaction</topic><topic>Promoter Regions, Genetic</topic><topic>Receptors, Thrombin - genetics</topic><topic>Restriction Mapping</topic><topic>Single-Strand Specific DNA and RNA Endonucleases - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, F</creatorcontrib><creatorcontrib>Baykal, D</creatorcontrib><creatorcontrib>Horaist, C</creatorcontrib><creatorcontrib>Yan, C N</creatorcontrib><creatorcontrib>Carr, B N</creatorcontrib><creatorcontrib>Rao, G N</creatorcontrib><creatorcontrib>Runge, M S</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, F</au><au>Baykal, D</au><au>Horaist, C</au><au>Yan, C N</au><au>Carr, B N</au><au>Rao, G N</au><au>Runge, M S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and Identification of Regulatory Sequences of the Human Thrombin Receptor Gene</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1996-10-18</date><risdate>1996</risdate><volume>271</volume><issue>42</issue><spage>26320</spage><epage>26328</epage><pages>26320-26328</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Thrombin, via activation of vascular endothelial and smooth muscle cell thrombin receptors, modulates vascular wall healing.
To understand the mechanisms that regulate human thrombin receptor (HTR) expression, we cloned and characterized the HTR gene.
The HTR gene consists of Exon I, which contains the 5â²-regulatory region and 85 nucleotides of coding sequence; a ~15-kb intron;
and Exon II, which contains the remainder of the coding sequence and the entire 3â²-untranslated region. Multiple transcription
initiation sites were identified by S1 mapping and ribonuclease protection assay. DNA sequence analysis indicated the presence
of two SP-1-AP-2 consensus binding sequences, near or within the transcription initiation sites, and consensus binding sequences
for numerous regulatory proteins that potentially modulate HTR expression. Functional analysis of the HTR promoter was performed
by transfecting human microvascular endothelial cells with HTR promoter region-luciferase constructs. The highest level of
expression was obtained with a 0.7-kb promoter sequence and was progressively less with fragments of 0.54, 1.16, 1.6, and
~3.2 kb. The data presented in this report provide a foundation for further characterization of the HTR gene and the mechanisms
that regulate its expression within the blood vessel wall.</abstract><cop>United States</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>8824285</pmid><doi>10.1074/jbc.271.42.26320</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1996-10, Vol.271 (42), p.26320-26328 |
| issn | 0021-9258 1083-351X |
| language | eng |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Amino Acid Sequence Base Sequence Blotting, Southern Chromosome Mapping Cloning, Molecular DNA Exons Gene Deletion Humans Introns Microscopy, Electron Molecular Sequence Data Polymerase Chain Reaction Promoter Regions, Genetic Receptors, Thrombin - genetics Restriction Mapping Single-Strand Specific DNA and RNA Endonucleases - metabolism |
| title | Cloning and Identification of Regulatory Sequences of the Human Thrombin Receptor Gene |
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