Identification of a neurorestrictive suppressor element (NRSE) in the human mu-opioid receptor gene.
Analysis of the DNA sequence of the human mu-opioid receptor gene (MOR) revealed that a region overlapping the start codon was substantially homologous to a DNA element named the neurorestrictive suppressor element (NRSE) or restrictive element 1 (RE-1). Transient transfection experiments in the L929 and HEK non-neural cell lines showed that expression of a MOR promoter/reporter gene construct was suppressed in non-neural cell lines by inclusion of this MOR NRSE. Expression from a thymidine kinase promoter was also suppressed when the MOR NRSE was inserted upstream or downstream of the reporter gene. The MOR NRSE did not suppress expression of the reporter gene in neural derived cell lines, IMR-32 and Neuro 2a. The transcription factor REST which binds NRSE thereby enacting the suppression of transcription, was encoded in a plasmid and co-transfected into the IMR-32 cells. The REST co-transfected neuronal derived (IMR-32) cells became sensitive to the MOR NRSE mediated suppression of reporter gene expression. Electrophoretic mobility shift experiments revealed that oligonucleotides containing the MOR NRSE were bound by a factor from nuclear extracts of non-neural cell lines, HeLa and Jurkat. This binding was specifically competed by oligonucleotides containing NRSE sequences previously shown to suppress transcription through REST. Thus an NRSE element overlapping the human MOR start codon suppresses gene expression in non-neural cell lines and may help direct neural tissue specific expression of MOR.
Andria
M L
ML
Department of Psychiatry, New York University Medical Center, 550 First Avenue, New York, NY10016, USA. adrim01@mcrcr.nyu.med.edu
Simon
E J
EJ
D020413:3' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D016384:Consensus Sequence; D019204:GTP-Binding Proteins / Q000502:physiology; D015870:Gene Expression / Q000502:physiology; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D017450:Receptors, Opioid, mu / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D014162:Transfection; D000099229:RE1-Silencing Transcription Factor
D020413:3' Untranslated Regions; D019204:GTP-Binding Proteins; D017450:Receptors, Opioid, mu; D012097:Repressor Proteins; D014157:Transcription Factors; D000099229:RE1-Silencing Transcription Factor