Identification of nuclear factor-kappaB responsive element within the neuronal nitric oxide synthase exon 1f-specific promoter.
Transcriptional regulation of the neuronal nitric oxide synthase gene (nNOS) is particularly complex as 12 distinct transcripts derived from different first exons are expressed in a tissue- and cell-specific manner. The exon 1f mRNA is relatively highly expressed in nervous system and relies upon exon 1f-specific promoter activity. Using conventional and real-time reverse transcription-polymerase chain reaction, we found exon 1f mRNA was the major transcript of the nNOS gene in human neuroblastoma SK-N-SH cells. We analyzed a 1090 bp fragment of 1f promoter by TRANSFAC-TESS and Match softwares and luciferase assay, and found an important positive transcriptional regulation region that contained a putative nuclear factor (NF)-kappaB binding site. Subsequently, using electrophoresis mobility shift and chromatin immunoprecipitation assays, we identified this site to be the NF-kappaB responsive element, a crucial positive regulator in the activation of the nNOS 1f promoter. Taken together, our study identified an NF-kappaB responsive element within nNOS 1f promoter and showed that it plays an important role in the transactivation of nNOS 1f mRNA, the major transcript of nNOS in SK-N-SH cells.
Li
Yinghui
Y
Department of Medical Genetics, China Medical University, Shenyang 110001, China.
Li
Guangyu
G
Li
Chunyi
C
Zhao
Yanyan
Y
D001483:Base Sequence; D001665:Binding Sites; D047369:Chromatin Immunoprecipitation; D024202:Electrophoretic Mobility Shift Assay; D005091:Exons; D006801:Humans; D016328:NF-kappa B / Q000235:genetics*; D009447:Neuroblastoma / Q000235:genetics*; D052248:Nitric Oxide Synthase Type I / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014158:Transcription, Genetic / Q000235:genetics*; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured
D016328:NF-kappa B; D052248:Nitric Oxide Synthase Type I