title: Role of testis-specific high-mobility-group protein in transcriptional regulation of inducible nitric oxide synthase expression in the liver of endotoxic shock mice.
abstract: Inducible nitric oxide synthase (iNOS) plays a central role in tissue damage during endotoxic shock. However, the underlying mechanisms that control transcription of iNOS are not completely defined. A screening strategy with DNA pull-down technology and two-dimensional difference in gel electrophorcsis (2D-DIGE) analysis was used to identify regulators that interact with the iNOS promoter. We found 14 proteins that bind to the iNOS promoter in the liver of endotoxic shock mice. After matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analysis, one of these proteins was identified as testis-specific high-mobility-group protein (tsHMG), an alternative splicing isoform encoded by the mitochondrial transcription factor A gene. We identified the binding sites of tsHMG on the iNOS promoter using a LiquiChip system, and further confirmed interactions between tsHMG and iNOS by RT-PCR, western blotting and immunofluorescence. Functional analysis by over-expression and RNA interference of tsHMG revealed that tsHMG regulates lipopolysaccharide-stimulated iNOS expression. These results indicate that tsHMG participates in modulation of iNOS expression in the early stage of endotoxic shock.
authors.affiliation: Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China; Department of Intensive Care Unit, General Hospital of Guangzhou Military Command, Guangzhou, China.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Liu
authors.forename: Jinghua
authors.initials: J
authors.lastname: Liu
authors.forename: Juan
authors.initials: J
authors.lastname: Wang
authors.forename: Jia
authors.initials: J
authors.lastname: Xu
authors.forename: Yawei
authors.initials: Y
authors.lastname: Liu
authors.forename: Xuegang
authors.initials: X
authors.lastname: Sun
authors.forename: Lei
authors.initials: L
authors.lastname: Su
authors.forename: Jiang Huai
authors.initials: JH
authors.lastname: Wang
authors.forename: Yong
authors.initials: Y
authors.lastname: Jiang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology*; D006609:High Mobility Group Proteins / Q000737:chemistry / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012772:Shock, Septic / Q000201:enzymology / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014158:Transcription, Genetic / Q000502:physiology*; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D008070:Lipopolysaccharides; C081930:Tfam protein, mouse; D052247:Nitric Oxide Synthase Type II
keywords: endotoxic shock; inducible nitric oxide synthase; lipopolysaccharide; testis-specific high-mobility-group protein; transcriptional regulation