Differential regulation of mouse and human Mu opioid receptor gene depends on the single stranded DNA structure of its promoter and α-complex protein 1

Lee,Dong‑Sun; Law,Ping‑Yee; Ln,Wei; Loh,Horace  H.; Song,Kyu  Young; Choi,Hack  Sun

doi:10.3892/br.2017.877

Differential regulation of mouse and human Mu opioid receptor gene depends on the single stranded DNA structure of its promoter and α-complex protein 1

Authors:
- Dong‑Sun Lee
- Ping‑Yee Law
- Wei Ln
- Horace H. Loh
- Kyu Young Song
- Hack Sun Choi
View Affiliations

Affiliations: Department of Biotechnology, College of Applied Life Science, Jeju National University, Jeju 63243, Republic of Korea, Department of Pharmacology, University of Minnesota Medical School, Minneapolis, MN 55455, USA
Published online on: March 21, 2017 https://doi.org/10.3892/br.2017.877
Pages: 532-538

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The Mu opioid receptor (MOR) mediates various functions of opioid-induced analgesia, euphoria and respiratory depression, and is a major target of opioid analgesics. Understanding of MOR gene expression among species is important for understanding its analgesic function in humans. In the current study, the polypyrimidine/polypurine (PPy/u) region, a key element of MOR gene expression, was compared in humans and mice. The mouse PPy/u element is highly homologous to its human element (84%), and the mouse MOR (mMOR) reporter drives luciferase activity 35‑fold more effectively than the human MOR (hMOR) reporter. The structural study of reporter plasmids using S1 nuclease indicates that the mouse PPy/u element has a particular conformational structure, namely a single‑stranded DNA (ssDNA) region that promotes strong promoter activity. DNA electrophoretic mobility shift assays demonstrated that the recombinant α‑complex protein 1 (α‑CP1) is capable of binding to a single‑stranded mouse PPy/u sequence. Furthermore, plasmid‑expressing α‑CP1 activated the expression of a luciferase reporter when cotransfected with a single‑stranded (p336/306) construct. In addition, the α‑CP1 gene induced the mMOR gene in mouse neuronal cells and did not induce the human neuronal MOR gene. The current study demonstrates that α‑CP1 functions as a transcriptional activator in the mMOR gene, but does not function in the hMOR gene due to species‑specific structural differences. The differences in human and mouse MOR gene expression are based on α‑CP1 and the ssDNA structure of the MOR promoter. The MOR gene is species‑specifically regulated, as the PPy/u element adopts a unique species-specific conformation and α-CP1 recruitment.

View Figures

View References

1	Min BH, Augustin LB, Felsheim RF, Fuchs JA and Loh HH: Genomic structure analysis of promoter sequence of a mouse mu opioid receptor gene. Proc Natl Acad Sci USA. 91:9081–9085. 1994. View Article : Google Scholar : PubMed/NCBI
2	Wei LN and Loh HH: Regulation of opioid receptor expression. Curr Opin Pharmacol. 2:69–75. 2002. View Article : Google Scholar : PubMed/NCBI
3	Kieffer BL: Recent advances in molecular recognition and signal transduction of active peptides: Receptors for opioid peptides. Cell Mol Neurobiol. 15:615–635. 1995. View Article : Google Scholar : PubMed/NCBI
4	Kieffer BL: Opioids: First lessons from knockout mice. Trends Pharmacol Sci. 20:19–26. 1999. View Article : Google Scholar : PubMed/NCBI
5	Law PY, Loh HH and Wei LN: Insights into the receptor transcription and signaling: Implications in opioid tolerance and dependence. Neuropharmacology. 47:(Suppl 1). S300–S311. 2004. View Article : Google Scholar
6	Law PY, Wong YH and Loh HH: Molecular mechanisms and regulation of opioid receptor signaling. Annu Rev Pharmacol Toxicol. 40:389–430. 2000. View Article : Google Scholar : PubMed/NCBI
7	Matthes HW, Maldonado R, Simonin F, Valverde O, Slowe S, Kitchen I, Befort K, Dierich A, Le Meur M, Dollé P, et al: Loss of morphine-induced analgesia, reward effect and withdrawal symptoms in mice lacking the mu-opioid-receptor gene. Nature. 383:819–823. 1996. View Article : Google Scholar : PubMed/NCBI
8	Mansour A, Fox CA, Akil H and Watson SJ: Opioid-receptor mRNA expression in the rat CNS: Anatomical and functional implications. Trends Neurosci. 18:22–29. 1995. View Article : Google Scholar : PubMed/NCBI
9	Uhl GR, Sora I and Wang Z: The mu opiate receptor as a candidate gene for pain: Polymorphisms, variations in expression, nociception and opiate responses. Proc Natl Acad Sci USA. 96:7752–7755. 1999. View Article : Google Scholar : PubMed/NCBI
10	Mogil JS: The genetic mediation of individual differences in sensitivity to pain and its inhibition. Proc Natl Acad Sci USA. 96:7744–7751. 1999. View Article : Google Scholar : PubMed/NCBI
11	Korostynski M, Kaminska-Chowaniec D, Piechota M and Przewlocki R: Gene expression profiling in the striatum of inbred mouse strains with distinct opioid-related phenotypes. BMC Genomics. 7:1462006. View Article : Google Scholar : PubMed/NCBI
12	Doyle GA, Sheng XR, Schwebel CL, Ferraro TN, Berrettini WH and Buono RJ: Identification and functional significance of polymorphisms in the mu-opioid receptor gene (Oprm) promoter of C57BL/6 and DBA/2 mice. Neurosci Res. 55:244–254. 2006. View Article : Google Scholar : PubMed/NCBI
13	Ko JL, Chen HC and Loh HH: Differential promoter usage of mouse mu-opioid receptor gene during development. Brain Res Mol Brain Res. 104:184–193. 2002. View Article : Google Scholar : PubMed/NCBI
14	Ko JL, Liu HC, Minnerath SR and Loh HH: Transcriptional regulation of mouse mu-opioid receptor gene. J Biol Chem. 273:27678–27685. 1998. View Article : Google Scholar : PubMed/NCBI
15	Ko JL and Loh HH: Single-stranded DNA-binding complex involved in transcriptional regulation of mouse mu-opioid receptor gene. J Biol Chem. 276:788–795. 2001. View Article : Google Scholar : PubMed/NCBI
16	Choe CY, Dong J, Law PY and Loh HH: Differential gene expression activity among species-specific polypyrimidine/polypurine motifs in mu opioid receptor gene promoters. Gene. 471:27–36. 2011. View Article : Google Scholar : PubMed/NCBI
17	Choi HS, Song KY, Hwang CK, Kim CS, Law PY, Wei LN and Loh HH: A proteomics approach for identification of single strand DNA-binding proteins involved in transcriptional regulation of mouse mu opioid receptor gene. Mol Cell Proteomics. 7:1517–1529. 2008. View Article : Google Scholar : PubMed/NCBI
18	Cook RJ, Karch C, Nahar P, Rivera A and Ko JL: Effects of desferoxamine-induced hypoxia on neuronal human mu-opioid receptor gene expression. Biochem Biophys Res Commun. 398:56–61. 2010. View Article : Google Scholar : PubMed/NCBI
19	Schroth GP and Ho PS: Occurrence of potential cruciform and H-DNA forming sequences in genomic DNA. Nucleic Acids Res. 23:1977–1983. 1995. View Article : Google Scholar : PubMed/NCBI
20	Kang DH, Song KY, Wei LN, Law PY, Loh HH and Choi HS: Novel function of the poly(c)-binding protein α-CP2 as a transcriptional activator that binds to single-stranded DNA sequences. Int J Mol Med. 32:1187–1194. 2013.PubMed/NCBI
21	Patterson SD and Aebersold R: Mass spectrometric approaches for the identification of gel-separated proteins. Electrophoresis. 16:1791–1814. 1995. View Article : Google Scholar : PubMed/NCBI
22	Hwang CK, Wu X, Wang G, Kim CS and Loh HH: Mouse mu opioid receptor distal promoter transcriptional regulation by SOX proteins. J Biol Chem. 278:3742–3750. 2003. View Article : Google Scholar : PubMed/NCBI
23	Choi HS, Hwang CK, Kim CS, Song KY, Law PY, Wei LN and Loh HH: Transcriptional regulation of mouse mu opioid receptor gene: Sp3 isoforms (M1, M2) function as repressors in neuronal cells to regulate the mu opioid receptor gene. Mol Pharmacol. 67:1674–1683. 2005. View Article : Google Scholar : PubMed/NCBI
24	Hardison RC: Comparative genomics. PLoS Biol. 1:E582003. View Article : Google Scholar : PubMed/NCBI
25	Wei LN and Loh HH: Transcriptional and epigenetic regulation of opioid receptor genes: Present and future. Annu Rev Pharmacol Toxicol. 51:75–97. 2011. View Article : Google Scholar : PubMed/NCBI
26	Vuillaumier S, Dixmeras I, Messaï H, Lapouméroulie C, Lallemand D, Gekas J, Chehab FF, Perret C, Elion J and Denamur E: Cross-species characterization of the promoter region of the cystic fibrosis transmembrane conductance regulator gene reveals multiple levels of regulation. Biochem J. 327:651–662. 1997. View Article : Google Scholar : PubMed/NCBI