Structure and Transcription of the Human m3 Muscarinic Receptor Gene

摘要: We have isolated and characterized the human m3 muscarinic receptor gene its promoter. Using 5' rapid amplification of cDNA ends (RACE), internal polymerase chain reaction (PCR), homology searching to identify EST clones, we determined that encoding comprises 4,559 bp in 8 exons, which are alternatively spliced exclude exons 2, 4, 6, and/or 7; coding sequence occurs within exon 8. Analysis P1 artificial chromosome (PAC) bacterial (BAC) clones PCR- amplified genomic DNA, 1 provided from Sanger Centre (Hinxton, Cambridge, UK) revealed spans at least 285 kb. A promoter fragment containing -1240 +101 (relative most transcription start site) exhibited considerable transcriptional activity during transient transfection cultured subconfluent, serum-fed canine tracheal myocytes, deletion analysis function presence positive regulatory elements between -526 -269. Sequence disclosed three potential AP-2 binding sites this region; five more consensus motifs occur -269 +101. Cotransfection with a plasmid expressing AP-2alpha substantially increased constructs 526 or 269 flanking DNA. Furthermore, was enhanced by long-term serum deprivation treatment is known increase transcription-promoting these cells. Together, data suggest expression regulated part airway smooth muscle.