Molecular Cloning and Functional Characterization of the Promoter Region of the Human Uncoupling Protein-2 Gene
Identifieur interne : 001529 ( Istex/Curation ); précédent : 001528; suivant : 001530Molecular 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.
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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<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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