δ-opioid receptor gene: Effect of Sp1 factor on transcriptional regulation in vivo

δ-Opioid receptor (DOR) promoter exhibited a cell-type-specific expression pattern. Protein-DNA interactions in this promoter were identified by dimethyl sulfate in vivo footprinting analysis of NG108-15 cells, expressing endogenous DOR. Complete protection of the putative Sp1 cis-element and partial protection of the sequence defined as X-Notl in the basal promoter were observed only in the G₀/G₁ phase of the cell cycle. No protection was detected in Neuro2A cells that do not express DOR. In vivo formaldehyde cross-linking confirmed Sp1 factor binding to its cis-acting element during the G₀/G₁ phase. The functional significance of these Sp1 and X-Notl sites was evaluated by transient transfection analysis. Northern blot analysis and nuclear run-off assays revealed maximum DOR mRNA level and transcription rate, respectively, during the G₀/G₁ phase of NG108-15 cells. In summary, the protein-DNA interactions at the Sp1 and X-Notl sites are necessary for cell cycle-dependent and cell-type-specific up-regulated DOR gene expression.