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Regulation of Mouse Neuropeptide Y Y 1 Receptor Gene Transcription: A Potential Role for Nuclear Factor-κB/Rel Proteins

Molecular Pharmacology, ISSN: 0026-895X, Vol: 51, Issue: 1, Page: 27-35
1997
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We previously isolated a 1.3-kb genomic fragment in the 5′-flanking region of the murine neuropeptide Y (NPY) Y 1 receptor gene, which is able to drive the expression of LacZ reporter gene in neuronal cells. We determined the ability of deletion mutants of this region to modulate transcription of the heterologous luciferase gene in the Y 1 receptor-expressing neuroblastoma/glioma NG108-15 cells and the Y 1 receptor-deficient 293 cells. Results suggest the presence of a cell type-specific core promoter (−399 to −218 from the initiator ATG) and, upstream, of two positive and two negative regulatory elements. Sequence analysis of the Y 1 receptor promoter identified two decameric sequences corresponding to consensus binding sites for nuclear factor-κB/Rel proteins. Gel shift analysis indicated that a 29-bp oligonucleotide comprising the two putative κB sites, which we refer to as Y 1 -κB sequence, specifically binds κB-related complexes in nuclear extracts from rat brain areas, NG108-15 cells, and the murine T cell clone A.E7. In nuclear extracts from A.E7 and NG108-15 cells, the Y 1 -κB sequence specifically binds an additional complex whose molecular nature remains to be elucidated. Through transient transfection studies, we also demonstrated that the Y 1 -κB sequence acts as an enhancer element, inferring its potential role in regulation of the Y 1 receptor gene expression.

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