Cloning and Functional Promoter Mapping of the Rat Serotonin-2 Receptor Gene

• Published in: Molecular and cellular neuroscience Vol. 5; no. 3; pp. 291 - 300
• Main Authors: Garlow, Steven J., Chin, Allison C., Marinovich, Adrian M., Heller, Mark R., Ciaranello, Roland D.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.06.1994
• Subjects: 3T3 Cells - metabolism; Animals; Base Sequence; Cells, Cultured; Cloning, Molecular; Cricetinae; Fibroblasts - metabolism; Genes; Mice; Molecular Sequence Data; Muscle, Smooth, Vascular - metabolism; Promoter Regions, Genetic; Rats - genetics; Receptors, Serotonin - classification; Receptors, Serotonin - genetics; Recombinant Fusion Proteins - biosynthesis; Sequence Alignment; Sequence Homology, Nucleic Acid; Species Specificity; Transcription, Genetic; Transfection
• ISSN: 1044-7431; 1095-9327
• DOI: 10.1006/mcne.1994.1034

We have cloned the gene encoding the rat serotonin-2 (5-HT 2) receptor. The transcription unit is divided into three exons by two introns. The major 5-HT 2 transcript is 5.62 kb in length and contains 1173 bases of 5′-untranslated region (5′-UTR) 1413 bases of open reading frame, and 3033 bases of 3′-UTR. Primer extension demonstrates one strong transcription initiation site 1173 nt from the start codon. Reverse transcriptase-polymerase chain reaction analysis indicates the presence of at least one additional minor site of transcription initiation 1355 nt from the start codon. There are ATA boxes 28 nt 5′ to both the major and minor sites of transcription initiation, Functional promoter mapping was carried out in a transient transfection assay. This analysis reveals that there are negative attenuating elements between 2.5 and 2.3 kb from the initiation site and positive elements between 1100 and 200 nt from transcription initiation. Minimal promoter sequences are contained within 200 nt of the major site of transcription initiation. These findings suggest that the expression of the 5-HT 2 receptor gene is regulated by a combination of positive and negative elements operating through the minimal promoter.