TY - JOUR T1 - δ-Opioid Receptor Gene: Effect of Sp1 Factor on Transcriptional Regulation in Vivo JF - Molecular Pharmacology JO - Mol Pharmacol SP - 331 LP - 340 DO - 10.1124/mol.60.2.331 VL - 60 IS - 2 AU - Dmitri Smirnov AU - Hee-Jeong Im AU - Horace H. Loh Y1 - 2001/08/01 UR - http://molpharm.aspetjournals.org/content/60/2/331.abstract N2 - 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-NotI 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-NotI 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-NotI sites are necessary for cell cycle-dependent and cell-type–specific up-regulated DOR gene expression. ER -