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Cloning and functional characterization of the human VIP1/PACAP receptor promoter
A Couvineau1, J J Maoret, C Rouyer-Fessard
1Laboratoire de Neuroendocrinologie et Biologie Cellulaire Digestives, Institut National de la Santé et de la Recherche Médicale, INSERM U-410, Faculté de Médecine Xavier Bichat, Paris, France. coucou@bichat.inserm.fr
Abstract:
The 5'-flanking region (1.5 kb) of the gene coding for the human VIP1/PACAP receptor was isolated, sequenced, and characterized. Transient expression of constructs containing sequentially deleted 5'-flanking sequences of the VIP1/PACAP receptor fused to a luciferase reporter gene showed that this sequence was active as a promoter in the intestinal cancer cell line, HT-29, expressing endogenous VIP1/PACAP receptor. The shortest DNA fragment with significant promoter activity encompassed the region from -205 to +76 bp. Deletion of a CCAAT-box sequence in the construction corresponding to -173 to +76 bp dramatically reduced the promoter activity. The promoter -205 to +76 bp has a housekeeping gene structure without TATA-box. It contains GC-rich regions characterized by potential Sp1 and AP2 sites and some potential regulatory elements, such as CRE and ATF, and a CCAAT-box sequence (-182 to -178) crucial for gene transcription.