Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene
Identifieur interne : 002477 ( Main/Merge ); précédent : 002476; suivant : 002478Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene
Auteurs : Naxin Tu [Allemagne] ; Hongmei Chen [Allemagne] ; Ulrike Winnikes [Allemagne] ; Irmtraud Reinert [Allemagne] ; Gabriele Marmann [Allemagne] ; Karl Martin Pirke [Allemagne] ; Klaus-Ulrich Lentes [Allemagne]Source :
- Biochemical and Biophysical Research Communications [ 0006-291X ] ; 1999.
English descriptors
- KwdEn :
- Animals, Base Sequence, Cell Line, Cloning, Molecular, DNA (genetics), DNA Primers (genetics), Gene Expression Regulation, Genes, Regulator, Humans, Ion Channels, Membrane Transport Proteins, Mitochondrial Proteins, Molecular Sequence Data, Promoter Regions, Genetic, Proteins (genetics), Recombinant Fusion Proteins (genetics), Transcription, Genetic, Uncoupling Agents, Uncoupling Protein 2.
- MESH :
- chemical , genetics : DNA, DNA Primers, Proteins, Recombinant Fusion Proteins.
- Animals, Base Sequence, Cell Line, Cloning, Molecular, Gene Expression Regulation, Genes, Regulator, Humans, Ion Channels, Membrane Transport Proteins, Mitochondrial Proteins, Molecular Sequence Data, Promoter Regions, Genetic, Transcription, Genetic, Uncoupling Agents, Uncoupling Protein 2.
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.
Url:
DOI: 10.1006/bbrc.1999.1663
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<front><div type="abstract" 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>
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<author><name sortKey="Chen, H" sort="Chen, H" uniqKey="Chen H" first="H" last="Chen">H. Chen</name>
</author>
<author><name sortKey="Winnikes, U" sort="Winnikes, U" uniqKey="Winnikes U" first="U" last="Winnikes">U. Winnikes</name>
</author>
<author><name sortKey="Reinert, I" sort="Reinert, I" uniqKey="Reinert I" first="I" last="Reinert">I. Reinert</name>
</author>
<author><name sortKey="Marmann, G" sort="Marmann, G" uniqKey="Marmann G" first="G" last="Marmann">G. Marmann</name>
</author>
<author><name sortKey="Pirke, K M" sort="Pirke, K M" uniqKey="Pirke K" first="K M" last="Pirke">K M Pirke</name>
</author>
<author><name sortKey="Lentes, K U" sort="Lentes, K U" uniqKey="Lentes K" first="K U" last="Lentes">K U Lentes</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="1999">1999</date>
<idno type="RBID">pubmed:10558866</idno>
<idno type="pmid">10558866</idno>
<idno type="doi">10.1006/bbrc.1999.1663</idno>
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<idno type="wicri:doubleKey">0006-291X:1999:Tu N:molecular:cloning:and</idno>
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<sourceDesc><biblStruct><analytic><title xml:lang="en">Molecular cloning and functional characterization of the promoter region of the human uncoupling protein-2 gene.</title>
<author><name sortKey="Tu, N" sort="Tu, N" uniqKey="Tu N" first="N" last="Tu">N. Tu</name>
<affiliation wicri:level="4"><nlm:affiliation>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, Trier, D-54290, Germany. tun@fpp.uni-trier.de</nlm:affiliation>
<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Laboratory of Molecular Neurogenetics, Center for Psychobiological and Psychosomatic Research (FPP), University of Trier, Trier, D-54290</wicri:regionArea>
<wicri:noRegion>54290</wicri:noRegion>
<orgName type="university">Université de Trèves</orgName>
<placeName><settlement type="city">Trèves (Allemagne)</settlement>
<region type="land" nuts="1">Rhénanie-Palatinat</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Chen, H" sort="Chen, H" uniqKey="Chen H" first="H" last="Chen">H. Chen</name>
</author>
<author><name sortKey="Winnikes, U" sort="Winnikes, U" uniqKey="Winnikes U" first="U" last="Winnikes">U. Winnikes</name>
</author>
<author><name sortKey="Reinert, I" sort="Reinert, I" uniqKey="Reinert I" first="I" last="Reinert">I. Reinert</name>
</author>
<author><name sortKey="Marmann, G" sort="Marmann, G" uniqKey="Marmann G" first="G" last="Marmann">G. Marmann</name>
</author>
<author><name sortKey="Pirke, K M" sort="Pirke, K M" uniqKey="Pirke K" first="K M" last="Pirke">K M Pirke</name>
</author>
<author><name sortKey="Lentes, K U" sort="Lentes, K U" uniqKey="Lentes K" first="K U" last="Lentes">K U Lentes</name>
</author>
</analytic>
<series><title level="j">Biochemical and biophysical research communications</title>
<idno type="ISSN">0006-291X</idno>
<imprint><date when="1999" type="published">1999</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Cell Line</term>
<term>Cloning, Molecular</term>
<term>DNA (genetics)</term>
<term>DNA Primers (genetics)</term>
<term>Gene Expression Regulation</term>
<term>Genes, Regulator</term>
<term>Humans</term>
<term>Ion Channels</term>
<term>Membrane Transport Proteins</term>
<term>Mitochondrial Proteins</term>
<term>Molecular Sequence Data</term>
<term>Promoter Regions, Genetic</term>
<term>Proteins (genetics)</term>
<term>Recombinant Fusion Proteins (genetics)</term>
<term>Transcription, Genetic</term>
<term>Uncoupling Agents</term>
<term>Uncoupling Protein 2</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA</term>
<term>DNA Primers</term>
<term>Proteins</term>
<term>Recombinant Fusion Proteins</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Cell Line</term>
<term>Cloning, Molecular</term>
<term>Gene Expression Regulation</term>
<term>Genes, Regulator</term>
<term>Humans</term>
<term>Ion Channels</term>
<term>Membrane Transport Proteins</term>
<term>Mitochondrial Proteins</term>
<term>Molecular Sequence Data</term>
<term>Promoter Regions, Genetic</term>
<term>Transcription, Genetic</term>
<term>Uncoupling Agents</term>
<term>Uncoupling Protein 2</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" 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>
</PubMed>
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