Studies of Dual Promoters of Mouse κ-Opioid Receptor Gene
Abstract
Dual promoters were identified in the mouse κ-opioid receptor (KOR) gene. The distal promoter was located in the 5′-upstream region of exon 1 and the proximal promoter was located in the first intron of this gene. The transcription initiation site of the proximal promoter was mapped to the −93rd nucleotide position from the ATG codon in a primer extension experiment. The expression of KOR mRNAs transcribed from these two promoters in mouse central nervous system and an embryonal carcinoma cell line P19 was confirmed in a ribonuclease protection assay. In non-neuronal tissues, only the transcripts initiated from the distal promoter were detected. The biological activities of these two promoters were determined in transient transfection of P19 cells with a series of reporters, each truncated at various 5′-upstream regions. It was concluded that the distal promoter was located between nucleotide positions −990 and −570, and the proximal promoter was located between nucleotide positions −330 and −93, relative to the translation initiation codon. The presence of dual promoters in the KOR gene suggested potential regulation of KOR expression by using different promoters.
Footnotes
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Send reprint requests to: Dr. Li-Na Wei, Department of Pharmacology, University of Minnesota Medical School, 3-249 Millard Hall, 435 Delaware Street S.E., Minneapolis, MN 55455. E-mail:weixx009{at}maroon.tc.umn.edu
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This study was supported by National Institutes of Health Grants DA01583, DA05695, K05-70554, and the F & A Stark Fund of Minnesota Medical Foundation (H.H.L.) and DK46866 and MMF-975-95 (L.N.W.)
- Abbreviations:
- KOR
- κ-opioid receptor
- TFDB
- transcription factor database
- RPA
- ribonuclease protection assay
- CNS
- central nervous system
- PCR
- polymerase chain reaction
- CHO
- Chinese hamster ovary
- RT
- reverse transcription
- bp
- base pair
- kb
- kilobase pair
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- Received February 20, 1996.
- Accepted May 8, 1997.
- The American Society for Pharmacology and Experimental Therapeutics
