,Identification of Nuclear Factor-κB Responsive Element within the Neuronal Nitric Oxide Synthase Ex

• 作 者: Yinghui LI Guangyu LI Chunyi LI Yanyan ZHAO
• 机 构: Department of Medical Genetics, China Medical University, Shenyang 110001, China“,”Department of Neu
• 出 处: 生物化学与生物物理学报（英文版）
• 发表日期: 2007年4期
• 关键词: nNOS NF-κB transactivation nervous system

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 cellspecific 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)-κB binding site. Subsequently, using electrophoresis mobility shift and chromatin immunoprecipitation assays, we identified this site to be the NF-κB responsive element, a crucial positive regulator in the activation of the nNOS 1f promoter. Taken together, our study identified an NF-κB 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.