title: Regulation of neuronal nitric oxide synthase exon 1f gene expression by nuclear factor-kappaB acetylation in human neuroblastoma cells.
abstract: The neuronal nitric oxide synthase (nNOS) is predominantly expressed in nervous tissues and subject to complex transcriptional controls. To determine the effect of acetylation on nNOS expression, human neuroblastoma SK-N-SH cells were treated with trichostatin A (TSA), a histone deacetylase inhibitor. As a consequence, total and exon 1f-specific nNOS mRNA, nNOS protein and nNOS-derived nitric oxide production were increased. Immunoprecipitation and western blot showed both nuclear factor-kappaB (NF-kappaB) subunits p65 and p50 were acetylated in the presence of TSA. The enhancement of the p65 and p50 acetylation was in accordance with their increased binding affinities to the NF-kappaB responsive element, which was identified at position -893 to -884 of the nNOS exon 1f promoter. Luciferase assays revealed that TSA up-regulated the transcriptional activity of the nNOS 1f promoter through NF-kappaB-mediated transactivation. Taken together, we demonstrate that acetylation plays a crucial role in nNOS expression and suggest that acetylation of NF-kappaB p65 and p50 subunits by TSA treatment may augment their DNA-binding affinities, thereby activating the nNOS exon 1f promoter. It may be one of the mechanisms by which acetylation modulates nNOS expression and nitric oxide output in SK-N-SH cells and may be the molecular basis for certain neurological disorders.
authors.affiliation: Department of Medical Genetics, China Medical University, Shenyang, China.
authors.forename: Yinghui
authors.initials: Y
authors.lastname: Li
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Guangyu
authors.initials: G
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Tingting
authors.initials: T
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Jingyu
authors.initials: J
authors.lastname: Lu
mesh_terms: D000107:Acetylation / Q000187:drug effects; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D005091:Exons; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007191:Indazoles / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D009447:Neuroblastoma; D052248:Nitric Oxide Synthase Type I / Q000235:genetics / Q000378:metabolism*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors; D014162:Transfection / Q000379:methods
chemical_list: D006877:Hydroxamic Acids; D007191:Indazoles; D016328:NF-kappa B; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; C012589:trichostatin A; D008156:Luciferases; D052248:Nitric Oxide Synthase Type I; C080122:7-nitroindazole