The potential promoter regions on the 5' flank sequence of the mu opioid receptor gene in lymphocytes.
The human mu opioid receptor is known to mediate a variety of physiological and pharmacological effects of morphine in many tissues. However, the molecular processes that regulate the expression of the mu opioid receptor gene in immune cells are not well understood. To study regulatory elements that affect the expression of the mu opioid receptor gene in human lymphocytes (LMOR), a 2,278 bp fragment of the 5' regulatory region of the mu opioid receptor gene was cloned and sequenced from CEM x174 cells. The transcriptional initiation site was mapped through a primer extension assay. A series of 5'-deleted plasmids were constructed and transiently transfected into cultured CEM x174 cells. The data indicated that morphine up-regulated the mRNA level of LMOR in a dose-dependent manner, which could be blocked by the opioid receptor antagonist naloxone. Only one transcription initiation site (TIS) about 110 bp upstream of the translation start codon was identified. The regions from -372 to -253 and -2279 to -1371 located in the 5' regulatory sequence of the mu opioid receptor gene contained enhancer elements, while the regions from -1371 to -968 and -650 to -370 possessed repressor elements. Those promoter elements were involved in the transcriptional regulation of the mu opioid receptor gene. Collectively, this data strongly indicates that the expression of the mu opioid receptor gene in lymphocytes is subject to the regulation of cis-elements upstream from the TIS.
Wei
Erman
E
Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100083, China.
Li
Pingfeng
P
Liu
Xinhua
X
Qian
Chaowei
C
Li
Hui
H
Xia
Weiyi
W
Li
Gang
G
D024506:5' Flanking Region / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017450:Receptors, Opioid, mu / Q000235:genetics*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000235:genetics
D017450:Receptors, Opioid, mu