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Functional characterization of the NF-κB binding site in the human NOD2 promoter

Cellular & Molecular Immunology volume 7, pages 288–295 (2010)Cite this article

Abstract

Nucleotide-binding and oligomerization domain 2 (NOD2), a member of the NOD protein family, plays an important role in innate immunity. In response to pathogen attack, NOD2 stimulates cytokine and defensin production by activating nuclear factor (NF)-κB, a key transcription factor responsible for mediating downstream reactions. However, the mechanism linking NOD2 regulation and NF-κB activation is poorly understood. Using bioinformatics, we found a completely preserved canonical NF-κB binding site in the NOD2 core promoter (−16 to −25 bp) in both humans and chimpanzees. The functional role of this NF-κB binding site was investigated using the enhanced green fluorescent protein (EGFP) reporter system, site-directed mutagenesis, the NF-κB activation inhibitor (JSH-23) and the chromatin immunoprecipitation (ChIP) assay. The results show that the NF-κB binding site is critical for regulation of the NOD2 gene. Either deletion of the NF-κB binding elements within the NOD2 promoter or treatment with an NF-κB activation inhibitor could lead to a significant loss of NOD2 promoter activity as detected by reporter gene assay. The canonical NF-κB binding site was bound by NF-κB as determined by the ChIP method. Based on these results, we suggest a positive feedback regulation between NF-κB and NOD2, which may represent an efficient mechanism in response to pathogen invasion.

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Acknowledgements

We thank Professor Yubing Zhou (Medical College of Jinan University) for her guidance and technical assistance. Our authors also appreciate the valuable discussion and amendment made by Dr Kum-heng Poon (Health Canada). This work was supported by grants from the Natural Science Foundation of Guangdong Province (No. 06025159) and the Natural Science Foundation from Department of Education of Guangdong Province (No. 126 (2005)).

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Author notes

  1. These authors contributed equally to this work.

  2. Dr SS Tang, Data Mining Department, Institute for Infocomm Research, 1 Fusionopolis Way, #21-01 Connexis, 138632, Singapore. E-mail: jdyhcf@sina.com

Authors and Affiliations

  1. Department of Pathophysiology, Medical College, Jinan University, Guangzhou, China

    Chaofeng Hu, Liping Sun, Daxiang Lu & Huadong Wang

  2. Department of Physiology and Biophysics, Faculty of Medicine, Dalhousie University, Halifax, NS, Canada

    Yiling Hu

  3. Data Mining Department, Institute for Infocomm Research, Singapore

    Suisheng Tang

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  1. Chaofeng Hu
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Hu, C., Sun, L., Hu, Y. et al. Functional characterization of the NF-κB binding site in the human NOD2 promoter. Cell Mol Immunol 7, 288–295 (2010). https://doi.org/10.1038/cmi.2010.16

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