Role of testis-specific high-mobility-group protein in transcriptional regulation of inducible nitric oxide synthase expression in the liver of endotoxic shock mice

Authors

  • Zhifeng Liu,

    1. Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China
    2. Department of Intensive Care Unit, General Hospital of Guangzhou Military Command, Guangzhou, China
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    • These authors contributed equally to this work
  • Jinghua Liu,

    1. Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China
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    • These authors contributed equally to this work
  • Juan Wang,

    1. Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China
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  • Jia Xu,

    1. Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China
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  • Yawei Liu,

    1. Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China
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  • Xuegang Sun,

    1. Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China
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  • Lei Su,

    1. Department of Intensive Care Unit, General Hospital of Guangzhou Military Command, Guangzhou, China
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  • Jiang Huai Wang,

    Corresponding author
    1. Department of Academic Surgery, University College Cork, Cork University Hospital, Ireland
    • Correspondence

      Y. Jiang, Department of Pathophysiology, Southern Medical University, Guangzhou 510515, China

      Fax: +86 20 87277521

      Tel: +86 20 61648231

      E-mail: yjiang@fimmu.com

      J. H. Wang, Department of Academic Surgery, University College Cork, Cork University Hospital, Cork, Ireland

      Fax: +353 21 4901240

      Tel: +353 21 4901275

      E-mail: jh.wang@ucc.ie

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  • Yong Jiang

    Corresponding author
    1. Key Laboratory of Functional Proteomics of Guangdong Province, Department of Pathophysiology, Southern Medical University, Guangzhou, China
    • Correspondence

      Y. Jiang, Department of Pathophysiology, Southern Medical University, Guangzhou 510515, China

      Fax: +86 20 87277521

      Tel: +86 20 61648231

      E-mail: yjiang@fimmu.com

      J. H. Wang, Department of Academic Surgery, University College Cork, Cork University Hospital, Cork, Ireland

      Fax: +353 21 4901240

      Tel: +353 21 4901275

      E-mail: jh.wang@ucc.ie

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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.

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