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Originally published In Press as doi:10.1074/jbc.M008923200 on December 11, 2000

J. Biol. Chem., Vol. 276, Issue 11, 7811-7819, March 16, 2001
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A New Mouse Liver-specific Gene, Encoding a Protein Homologous to Human Antimicrobial Peptide Hepcidin, Is Overexpressed during Iron Overload*

Christelle PigeonDagger §, Gennady IlyinDagger , Brice CourselaudDagger , Patricia LeroyerDagger , Bruno TurlinDagger , Pierre BrissotDagger ||, and Olivier LoréalDagger

From the Dagger  INSERM U522, CHRU Pontchaillou, Rennes,  Laboratoire d'Anatomo-Pathologie B, CHRU Pontchaillou, Rennes, and || Service des Maladies du Foie, CHRU Pontchaillou, 35033 Rennes, France

Considering that the development of hepatic lesions related to iron overload diseases might be a result of abnormally expressed hepatic genes, we searched for new genes up-regulated under the condition of iron excess. By suppressive subtractive hybridization performed between livers from carbonyl iron-overloaded and control mice, we isolated a 225-base pair cDNA. By Northern blot analysis, the corresponding mRNA was confirmed to be overexpressed in livers of experimentally (carbonyl iron and iron-dextran-treated mice) and spontaneously (beta 2-microglobulin knockout mice) iron-overloaded mice. In addition, beta 2-microglobulin knockout mice fed with a low iron content diet exhibited a decrease of hepatic mRNA expression. The murine full-length cDNA was isolated and was found to encode an 83-amino acid protein presenting a strong homology in its C-terminal region to the human antimicrobial peptide hepcidin. In addition, we cloned the corresponding rat and human orthologue cDNAs. Both mouse and human genes named HEPC are constituted of 3 exons and 2 introns and are located on chromosome 7 and 19, respectively, in close proximity to USF2 gene. In mouse and human, HEPC mRNA was predominantly expressed in the liver. During both in vivo and in vitro studies, HEPC mRNA expression was enhanced in mouse hepatocytes under the effect of lipopolysaccharide. Finally, to analyze the intracellular localization of the predicted protein, we used the green fluorescent protein chimera expression vectors. The murine green fluorescent protein-prohepcidin protein was exclusively localized in the nucleus. When the putative nuclear localization signal was deleted, the resulting protein was addressed to the cytoplasm. Taken together, our data strongly suggest that the product of the new liver-specific gene HEPC might play a specific role during iron overload and exhibit additional functions distinct from its antimicrobial activity.

* This work was supported by La Ligue Contre le Cancer (Comité d'Ille et Vilaine), BIOMED 2 Grant CE BMH4-CT97-2149, the Ministère de la Recherche et de la Technologie, l'Association pour la Recherche Contre le Cancer, and l'Association Fer et Foie.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF297664, AF309489, and AF344185.

§ To whom correspondence should be addressed: INSERM U522, CHRU Pontchaillou, 35033 Rennes Cedex, France. Tel.: 33 299543737; Fax: 33 299540137; E-mail: christelle.pigeon@rennes.inserm.fr.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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