Cloning and functional promoter mapping of the rat serotonin-2 receptor gene.
We have cloned the gene encoding the rat serotonin-2 (5-HT2) receptor. The transcription unit is divided into three exons by two introns. The major 5-HT2 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-HT2 receptor gene is regulated by a combination of positive and negative elements operating through the minimal promoter.
Garlow
S J
SJ
Nancy Pritzker Laboratory of Developmental and Molecular Neurobiology, Department of Psychiatry and Behavioral Sciences, Stanford University, California 94305-5485.
Chin
A C
AC
Marinovich
A M
AM
Heller
M R
MR
Ciaranello
R D
RD
D016475:3T3 Cells / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D005347:Fibroblasts / Q000378:metabolism; D005796:Genes; D051379:Mice; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051381:Rats / Q000235:genetics*; D011985:Receptors, Serotonin / Q000145:classification / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014158:Transcription, Genetic; D014162:Transfection
D011985:Receptors, Serotonin; D011993:Recombinant Fusion Proteins