A major species of mouse mu-opioid receptor mRNA and its promoter-dependent functional polyadenylation signal.
The pharmacological effects of opioid drugs are mediated mainly by the mu-opioid receptor (MOR), which is encoded by an mRNA transcript named MOR1. Although several MOR mRNA splice variants have been reported, their biological relevance has been debated. In this study, we found that probes of regions essential for the production of functional MOR, as well as that of the 3'-downstream region of the MOR gene coding region, detected by Northern blot analyses, a major species of mature transcript MOR1 from mouse brain of approximately 11.5 kilobases (kb). Although exon 3 probe detected an additional 3.7-kb transcript, this transcript was not detected by other probes, ruling out its ability to produce functional MOR. The 3'-untranslated region (UTR) of MOR1 is contiguously extended from the end of the coding region, and uses a single polyadenylation [poly (A)] signal (located 10,179 bp downstream of the MOR1 stop codon). The poly (A) signal (AAUAAA) is located 26 bp upstream of the poly (A) site. Transient transfection using luciferase reporters verified the functionality of this poly (A) signal, in particular on a reporter driven by the MOR promoter. This poly (A) is much less effective for a heterologous promoter, such as simian virus 40, indicating a functional coupling of MOR promoter and its own poly (A). This report verifies MOR1 as the major mature MOR gene transcript that has the full capacity to produce functional MOR protein, identifies the 3'-UTR of MOR1 transcript, and uncovers functional coupling of the MOR gene promoter and its polyadenylation signal.
Wu
Qifang
Q
Department of Pharmacology, University of Minnesota Medical School, 6-120 Jackson Hall, 321 Church St. S.E., Minneapolis, MN 55455, USA. wuxx0285@umn.edu
Hwang
Cheol Kyu
CK
Yao
Shiyi
S
Law
Ping-Yee
PY
Loh
Horace H
HH
Wei
Li-Na
LN
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D039104:RNA 3' Polyadenylation Signals / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics*; D017450:Receptors, Opioid, mu / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000502:physiology
D012333:RNA, Messenger; D017450:Receptors, Opioid, mu