Transcription of the rat cholecystokinin gene is initiated at multiple sites: verification by an in vitro transcription system.

The cDNA that accommodates the most distal 5'-end region of cholecystokinin mRNA was isolated from an internally primed cDNA library. Using primer extension and S1 nuclease protection analyses, we demonstrated multiple RNA molecules generated from the rat cholecystokinin gene, a single-copy sequence. The longest RNA is transcribed at position--225 upstream relative to the translation start site. The major transcription, more than 95% of the total cholecystokinin mRNA in rat brain, occurred at--59 and its promoter activity was determined by in vitro RNA synthesis in a HeLa cell extract. Deletion to--105 demonstrated an approximately 60% decrease in transcriptional level compared with the full promoter activity. At least the upstream region between--254 and--105 is necessary for transcription initiated at--59 of the cholecystokinin gene by the cell-free system.