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Friend of GATA and GATA-6 modulate the transcriptional up-regulation of hepcidin in hepatocytes during inflammation

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Abstract

Hepcidin is an antimicrobial peptide hormone that plays a central role in the metabolism of iron and its expression in the liver can be induced through two major pathways: the inflammatory pathway, mainly via IL-6; and the iron-sensing pathway, mediated by BMP-6. GATA-proteins are group of evolutionary conserved transcriptional regulators that bind to the consensus motif—WGATAR—in the promoter region. In hepatoma cells, GATA-proteins 4 and 6 in conjunction with the co-factor friend of GATA (FOG) were shown to modulate the transcription of HAMP. However, it is unclear as to which of the GATA-proteins drive the expression of HAMP in vivo. In this study, using in vitro and in vivo approaches, we investigated the relevance of GATA and FOG proteins in the expression of hepcidin following treatment with IL-6 and BMP-6. We found that treatment of Huh7 cells with either IL-6 or BMP-6 increased the HAMP promoter activity. The HAMP promoter activity following treatment with IL-6 or BMP-6 was further increased by co-transfection of the promoter with GATA proteins 4 and 6. However, co-transfection of the HAMP promoter with FOG proteins 1 or 2 repressed the promoter response to treatments with either IL-6 or BMP-6. The effects of both GATA and FOG proteins on the promoter activity in response to IL-6 or BMP-6 treatment were abrogated by mutation of the GATA response element—TTATCT—in the HAMP promoter region −103/−98. In vivo, treatment of mice with lipopolysaccharide led to a transient increase of Gata-6 expression in the liver that was positively correlated with the expression of hepcidin. Our results indicate that during inflammation GATA-6 is up-regulated in concert with hepcidin while GATA-4 and FOG (1 and 2) are repressed.

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Acknowledgments

We are grateful to Dr Robert Viger for the GATA (1, 3, 4, and 6) and FOG (1 and 2) expression vectors, and Dr Jacques Tremblay for the pXP1 luciferase reporter plasmid. We also thank Drs Muhammad Zafarullah and Richard Betrand (CRCHUM) for their technical help and advice.

Disclosure

The authors declare there is no conflict of interest that would prejudice their impartiality. ETB developed, designed and executed the experiments, collected, analyzed and interpreted the data and then wrote the manuscript. AL performed the animal experiments. MMS participated in the data discussions and writing of the manuscript.

Funding

This work was supported by grants from the Canadian Institutes of Health Research (CIHR, Grant No. MOP123246) and Natural Sciences and Engineering Research Council of Canada (NSERC, Grant No. 412326-2011) to MMS. ETB is a recipient of Post-doctoral fellowship grants from the Institut du Cancer de Montréal, CHUM and the Centre de Recherche du CHUM, Université de Montréal QC, Canada. AL is a recipient of a PhD scholarship from the NSERC. MMS is a recipient of the Research Scholarship (Senior) from Fonds de recherche du Québec—Santé (FRQS).

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Authors and Affiliations

  1. Centre de Recherche du Centre Hospitalier de l’Université de Montréal (CRCHUM), Institut du Cancer de Montréal, (ICM), University of Montreal, Pavillon De Sève Porte Y-5625, 2099 rue Alexandre De Sève, Montreal, QC, H2L 4M1, Canada

    Edward T. Bagu, A. Layoun, A. Calvé & M. M. Santos

  2. Departement of Medicine, University of Montreal, Montreal, QC, Canada

    M. M. Santos

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  1. Edward T. Bagu
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  2. A. Layoun
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Correspondence to Edward T. Bagu.

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10534_2013_9683_MOESM1_ESM.pptx

Supplementary material 1 The fold change in mRNA level of both hepcidin in the mouse liver following treatment with LPS. Hepatic mRNA levels of hepcidin in wild-type (Wt) mice were determined 1.5, 3, 6, 12 and 24 h after treatment with LPS. Mice in the control group (time = 0) were injected a sterile saline solution. The data is presented as mean ± SEM with n = 4 mice/group. Results of each time point are representative of three independent experiments performed. NS, ** (P ≤ 0.05); *(< 0.01) are used in reference to post treatments groups 1.5, 3, and 6 h respectively when compared with the control group (0 h). (PPTX 48 kb)

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Bagu, E.T., Layoun, A., Calvé, A. et al. Friend of GATA and GATA-6 modulate the transcriptional up-regulation of hepcidin in hepatocytes during inflammation. Biometals 26, 1051–1065 (2013). https://doi.org/10.1007/s10534-013-9683-6

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  • DOI: https://doi.org/10.1007/s10534-013-9683-6

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