Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene
Identifieur interne : 001641 ( Istex/Corpus ); précédent : 001640; suivant : 001642Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene
Auteurs : Naxin Tu ; Hongmei Chen ; Ulrike Winnikes ; Irmtraud Reinert ; Gabriele Marmann ; Karl Martin Pirke ; Klaus-Ulrich LentesSource :
- Biochemical and Biophysical Research Communications [ 0006-291X ] ; 1999.
Abstract
As a member of the uncoupling protein family, UCP2 is ubiquitously expressed in rodents and humans, implicating a major role in thermogenesis. To analyze promoter function and regulatory motifs involved in the transcriptional regulation of UCP2 gene expression, 3.3 kb of 5′-flanking region of the human UCP2 (hUCP2) gene have been cloned. Sequence analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box, however, appeared GC-rich resulting in the presence of several Sp-1 motifs and Ap-1/-2 binding sites near the transcription initiation site. Functional characterization of human UCP2 promoter-CAT fusion constructs in transient expression assays showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site (+1). 75 bp further upstream (from nt −141 to −66) a strong cis-acting regulatory element (or enhancer) was identified, which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements distributed over a minimum of 3.3 kb of the promoter region.
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DOI: 10.1006/bbrc.1999.1663
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xml:lang="en">As a member of the uncoupling protein family, UCP2 is ubiquitously expressed in rodents and humans, implicating a major role in thermogenesis. To analyze promoter function and regulatory motifs involved in the transcriptional regulation of UCP2 gene expression, 3.3 kb of 5′-flanking region of the human UCP2 (hUCP2) gene have been cloned. Sequence analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box, however, appeared GC-rich resulting in the presence of several Sp-1 motifs and Ap-1/-2 binding sites near the transcription initiation site. Functional characterization of human UCP2 promoter-CAT fusion constructs in transient expression assays showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site (+1). 75 bp further upstream (from nt −141 to −66) a strong cis-acting regulatory element (or enhancer) was identified, which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements distributed over a minimum of 3.3 kb of the promoter region.</div></front></TEI><istex><corpusName>elsevier</corpusName><author><json:item><name>Naxin Tu</name><affiliations><json:string>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</json:string></affiliations></json:item><json:item><name>Hongmei Chen</name><affiliations><json:string>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</json:string></affiliations></json:item><json:item><name>Ulrike Winnikes</name><affiliations><json:string>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</json:string></affiliations></json:item><json:item><name>Irmtraud Reinert</name><affiliations><json:string>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</json:string></affiliations></json:item><json:item><name>Gabriele Marmann</name><affiliations><json:string>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</json:string></affiliations></json:item><json:item><name>Karl Martin Pirke</name><affiliations><json:string>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</json:string></affiliations></json:item><json:item><name>Klaus-Ulrich Lentes</name><affiliations><json:string>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</json:string></affiliations></json:item></author><articleId><json:string>91663</json:string></articleId><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>As a member of the uncoupling protein family, UCP2 is ubiquitously expressed in rodents and humans, implicating a major role in thermogenesis. To analyze promoter function and regulatory motifs involved in the transcriptional regulation of UCP2 gene expression, 3.3 kb of 5′-flanking region of the human UCP2 (hUCP2) gene have been cloned. Sequence analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box, however, appeared GC-rich resulting in the presence of several Sp-1 motifs and Ap-1/-2 binding sites near the transcription initiation site. Functional characterization of human UCP2 promoter-CAT fusion constructs in transient expression assays showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site (+1). 75 bp further upstream (from nt −141 to −66) a strong cis-acting regulatory element (or enhancer) was identified, which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements distributed over a minimum of 3.3 kb of the promoter region.</abstract><qualityIndicators><score>7.016</score><pdfVersion>1.3</pdfVersion><pdfPageSize>551 x 744 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>0</keywordCount><abstractCharCount>1144</abstractCharCount><pdfWordCount>5131</pdfWordCount><pdfCharCount>30752</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>168</abstractWordCount></qualityIndicators><title>Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene</title><pii><json:string>S0006-291X(99)91663-7</json:string></pii><refBibs><json:item><host><volume>64</volume><pages><last>64</last><first>1</first></pages><author></author><title>Physiol. 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CAT, chloramphenicol acetyltransferase; ELISA, enzyme-linked immunosorbent assay.</note><note type="content">Section title: Regular Article</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene</title><author xml:id="author-1"><persName><forename type="first">Naxin</forename><surname>Tu</surname></persName><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation></author><author xml:id="author-2"><persName><forename type="first">Hongmei</forename><surname>Chen</surname></persName><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation></author><author xml:id="author-3"><persName><forename type="first">Ulrike</forename><surname>Winnikes</surname></persName><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation></author><author xml:id="author-4"><persName><forename type="first">Irmtraud</forename><surname>Reinert</surname></persName><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation></author><author xml:id="author-5"><persName><forename type="first">Gabriele</forename><surname>Marmann</surname></persName><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation></author><author xml:id="author-6"><persName><forename type="first">Karl Martin</forename><surname>Pirke</surname></persName><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation></author><author xml:id="author-7"><persName><forename type="first">Klaus-Ulrich</forename><surname>Lentes</surname></persName><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation></author></analytic><monogr><title level="j">Biochemical and Biophysical Research Communications</title><title level="j" type="abbrev">YBBRC</title><idno type="pISSN">0006-291X</idno><idno type="PII">S0006-291X(00)X0061-7</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1999"></date><biblScope unit="volume">265</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="326">326</biblScope><biblScope unit="page" to="334">334</biblScope></imprint></monogr><idno type="istex">B47DB1D0077E7AB8546F3487D18FE11CFA0846F8</idno><idno type="DOI">10.1006/bbrc.1999.1663</idno><idno type="PII">S0006-291X(99)91663-7</idno><idno type="ArticleID">91663</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1999</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>As a member of the uncoupling protein family, UCP2 is ubiquitously expressed in rodents and humans, implicating a major role in thermogenesis. To analyze promoter function and regulatory motifs involved in the transcriptional regulation of UCP2 gene expression, 3.3 kb of 5′-flanking region of the human UCP2 (hUCP2) gene have been cloned. Sequence analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box, however, appeared GC-rich resulting in the presence of several Sp-1 motifs and Ap-1/-2 binding sites near the transcription initiation site. Functional characterization of human UCP2 promoter-CAT fusion constructs in transient expression assays showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site (+1). 75 bp further upstream (from nt −141 to −66) a strong cis-acting regulatory element (or enhancer) was identified, which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements distributed over a minimum of 3.3 kb of the promoter region.</p></abstract></profileDesc><revisionDesc><change when="1999">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/document/B47DB1D0077E7AB8546F3487D18FE11CFA0846F8/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="UTF-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla" xml:lang="en"><item-info><jid>YBBRC</jid><aid>91663</aid><ce:pii>S0006-291X(99)91663-7</ce:pii><ce:doi>10.1006/bbrc.1999.1663</ce:doi><ce:copyright type="full-transfer" year="1999">Academic Press</ce:copyright></item-info><head><ce:article-footnote><ce:label>☆</ce:label><ce:note-para>Abbreviations used: UCP, uncoupling protein; CAT, chloramphenicol acetyltransferase; ELISA, enzyme-linked immunosorbent assay.</ce:note-para></ce:article-footnote><ce:dochead><ce:textfn>Regular Article</ce:textfn></ce:dochead><ce:title>Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene</ce:title><ce:author-group><ce:author><ce:given-name>Naxin</ce:given-name><ce:surname>Tu</ce:surname><ce:cross-ref refid="FN1"><ce:sup loc="post">1</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Hongmei</ce:given-name><ce:surname>Chen</ce:surname></ce:author><ce:author><ce:given-name>Ulrike</ce:given-name><ce:surname>Winnikes</ce:surname></ce:author><ce:author><ce:given-name>Irmtraud</ce:given-name><ce:surname>Reinert</ce:surname></ce:author><ce:author><ce:given-name>Gabriele</ce:given-name><ce:surname>Marmann</ce:surname></ce:author><ce:author><ce:given-name>Karl Martin</ce:given-name><ce:surname>Pirke</ce:surname></ce:author><ce:author><ce:given-name>Klaus-Ulrich</ce:given-name><ce:surname>Lentes</ce:surname></ce:author><ce:affiliation><ce:textfn>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</ce:textfn></ce:affiliation><ce:footnote id="FN1"><ce:label>1</ce:label><ce:note-para>To whom correspondence should be addressed. Fax: 49-651-97504-90. E-mail: tun@fpp.uni-trier.de.</ce:note-para></ce:footnote></ce:author-group><ce:date-received day="27" month="9" year="1999"></ce:date-received><ce:abstract class="author"><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para view="all" id="simple-para.0010">As a member of the uncoupling protein family, UCP2 is ubiquitously expressed in rodents and humans, implicating a major role in thermogenesis. To analyze promoter function and regulatory motifs involved in the transcriptional regulation of UCP2 gene expression, 3.3 kb of 5′-flanking region of the human UCP2 (hUCP2) gene have been cloned. Sequence analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box, however, appeared GC-rich resulting in the presence of several Sp-1 motifs and Ap-1/-2 binding sites near the transcription initiation site. Functional characterization of human UCP2 promoter-CAT fusion constructs in transient expression assays showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site (+1). 75 bp further upstream (from nt −141 to −66) a strong <ce:italic>cis</ce:italic>-acting regulatory element (or enhancer) was identified, which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements distributed over a minimum of 3.3 kb of the promoter region.</ce:simple-para></ce:abstract-sec></ce:abstract></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene</title></titleInfo><name type="personal"><namePart type="given">Naxin</namePart><namePart type="family">Tu</namePart><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation><description>To whom correspondence should be addressed. Fax: 49-651-97504-90. E-mail: tun@fpp.uni-trier.de.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Hongmei</namePart><namePart type="family">Chen</namePart><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ulrike</namePart><namePart type="family">Winnikes</namePart><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Irmtraud</namePart><namePart type="family">Reinert</namePart><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Gabriele</namePart><namePart type="family">Marmann</namePart><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Karl Martin</namePart><namePart type="family">Pirke</namePart><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Klaus-Ulrich</namePart><namePart type="family">Lentes</namePart><affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, D-54290, Trier, Germany</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article"></genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1999</dateIssued><copyrightDate encoding="w3cdtf">1999</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">As a member of the uncoupling protein family, UCP2 is ubiquitously expressed in rodents and humans, implicating a major role in thermogenesis. To analyze promoter function and regulatory motifs involved in the transcriptional regulation of UCP2 gene expression, 3.3 kb of 5′-flanking region of the human UCP2 (hUCP2) gene have been cloned. Sequence analysis showed that the promoter region of hUCP2 lacks a classical TATA or CAAT box, however, appeared GC-rich resulting in the presence of several Sp-1 motifs and Ap-1/-2 binding sites near the transcription initiation site. Functional characterization of human UCP2 promoter-CAT fusion constructs in transient expression assays showed that minimal promoter activity was observed within 65 bp upstream of the transcriptional start site (+1). 75 bp further upstream (from nt −141 to −66) a strong cis-acting regulatory element (or enhancer) was identified, which significantly enhanced basal promoter activity. The regulation of human UCP2 gene expression involves complex interactions among positive and negative regulatory elements distributed over a minimum of 3.3 kb of the promoter region.</abstract><note>Abbreviations used: UCP, uncoupling protein; CAT, chloramphenicol acetyltransferase; ELISA, enzyme-linked immunosorbent assay.</note><note type="content">Section title: Regular Article</note><relatedItem type="host"><titleInfo><title>Biochemical and Biophysical Research Communications</title></titleInfo><titleInfo type="abbreviated"><title>YBBRC</title></titleInfo><genre type="journal">journal</genre><originInfo><dateIssued encoding="w3cdtf">19991119</dateIssued></originInfo><identifier type="ISSN">0006-291X</identifier><identifier type="PII">S0006-291X(00)X0061-7</identifier><part><date>19991119</date><detail type="volume"><number>265</number><caption>vol.</caption></detail><detail type="issue"><number>2</number><caption>no.</caption></detail><extent unit="issue pages"><start>273</start><end>610</end></extent><extent unit="pages"><start>326</start><end>334</end></extent></part></relatedItem><identifier type="istex">B47DB1D0077E7AB8546F3487D18FE11CFA0846F8</identifier><identifier type="DOI">10.1006/bbrc.1999.1663</identifier><identifier type="PII">S0006-291X(99)91663-7</identifier><identifier type="ArticleID">91663</identifier><accessCondition type="use and reproduction" contentType="copyright">©1999 Academic Press</accessCondition><recordInfo><recordContentSource>ELSEVIER</recordContentSource><recordOrigin>Academic Press, ©1999</recordOrigin></recordInfo></mods></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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