Journal of Molecular Medicine

, Volume 87, Issue 5, pp 471–480

Bone morphogenetic protein (BMP)-responsive elements located in the proximal and distal hepcidin promoter are critical for its response to HJV/BMP/SMAD

  • Guillem Casanovas
  • Katarzyna Mleczko-Sanecka
  • Sandro Altamura
  • Matthias W. Hentze
  • Martina U. Muckenthaler
Original Article

Abstract

The hemochromatosis proteins HFE, transferrin receptor 2 (TfR2) and hemojuvelin (HJV, HFE2) positively control expression of the major iron regulatory hormone hepcidin. HJV is a bone morphogenetic protein (BMP) co-receptor that enhances the cellular response to BMP cytokines via the phosphorylation of SMAD proteins. In this study, we show that two highly conserved and sequence-identical BMP-responsive elements located at positions −84/−79 (BMP-RE1) and −2,255/−2,250 (BMP-RE2) of the human hepcidin promoter are critical for both the basal hepcidin mRNA expression and the hepcidin response to BMP-2 and BMP-6. While BMP-RE1 and BMP-RE2 show additive effects in responding to HJV-mediated BMP signals, only BMP-RE1 that is located in close proximity to a previously identified STAT-binding site is important for the hepcidin response to IL-6. These data identify a missing link between the HJV/BMP signaling pathways and hepcidin transcription, and further define the connection between inflammation and BMP-dependent hepcidin promoter activation. As such, they provide important new information furthering our understanding of disorders of iron metabolism and the anemia of inflammation.

Keywords

Hereditary hemochromatosis Hemojuvelin SMAD Anemia of inflammation IL-6 

Supplementary material

109_2009_447_MOESM1_ESM.ppt (138 kb)
Supplementary Table 1Primer pairs used for site-directed mutagenesis to generate luciferase reporter vectors (PPT 137 KB).

References

  1. 1.
    Andrews NC (2008) Forging a field: the golden age of iron biology. Blood 112:219–230CrossRefPubMedGoogle Scholar
  2. 2.
    Nemeth E, Tuttle MS, Powelson J, Vaughn MB, Donovan A, Ward DM, Ganz T, Kaplan J (2004) Hepcidin regulates cellular iron efflux by binding to ferroportin and inducing its internalization. Science 306:2090–2093CrossRefPubMedGoogle Scholar
  3. 3.
    Papanikolaou G, Samuels ME, Ludwig EH, MacDonald ML, Franchini PL, Dube MP, Andres L, MacFarlane J, Sakellaropoulos N, Politou M, Nemeth E, Thompson J, Risler JK, Zaborowska C, Babakaiff R, Radomski CC, Pape TD, Davidas O, Christakis J, Brissot P, Lockitch G, Ganz T, Hayden MR, Goldberg YP (2004) Mutations in HFE2 cause iron overload in chromosome 1q-linked juvenile hemochromatosis. Nat Genet 36:77–82CrossRefPubMedGoogle Scholar
  4. 4.
    Bridle KR, Frazer DM, Wilkins SJ, Dixon JL, Purdie DM, Crawford DH, Subramaniam VN, Powell LW, Anderson GJ, Ramm GA (2003) Disrupted hepcidin regulation in HFE-associated haemochromatosis and the liver as a regulator of body iron homoeostasis. Lancet 361:669–673CrossRefPubMedGoogle Scholar
  5. 5.
    Nemeth E, Roetto A, Garozzo G, Ganz T, Camaschella C (2005) Hepcidin is decreased in TFR2 hemochromatosis. Blood 105:1803–1806CrossRefPubMedGoogle Scholar
  6. 6.
    Verga Falzacappa MV, Muckenthaler MU (2005) Hepcidin: iron-hormone and anti-microbial peptide. Gene 364:37–44CrossRefPubMedGoogle Scholar
  7. 7.
    Babitt JL, Huang FW, Wrighting DM, Xia Y, Sidis Y, Samad TA, Campagna JA, Chung RT, Schneyer AL, Woolf CJ, Andrews NC, Lin HY (2006) Bone morphogenetic protein signaling by hemojuvelin regulates hepcidin expression. Nat Genet 38:531–539CrossRefPubMedGoogle Scholar
  8. 8.
    Shi Y, Massague J (2003) Mechanisms of TGF-beta signaling from cell membrane to the nucleus. Cell 113:685–700CrossRefPubMedGoogle Scholar
  9. 9.
    Wang RH, Li C, Xu X, Zheng Y, Xiao C, Zerfas P, Cooperman S, Eckhaus M, Rouault T, Mishra L, Deng CX (2005) A role of SMAD4 in iron metabolism through the positive regulation of hepcidin expression. Cell Metab 2:399–409CrossRefPubMedGoogle Scholar
  10. 10.
    Verga Falzacappa MV, Casanovas G, Hentze MW, Muckenthaler MU (2008) A bone morphogenetic protein (BMP)-responsive element in the hepcidin promoter controls HFE2-mediated hepatic hepcidin expression and its response to IL-6 in cultured cells. J Mol Med 86:531–540CrossRefPubMedGoogle Scholar
  11. 11.
    Verga Falzacappa MV, Vujic Spasic M, Kessler R, Stolte J, Hentze MW, Muckenthaler MU (2007) STAT3 mediates hepatic hepcidin expression and its inflammatory stimulation. Blood 109:353–358CrossRefPubMedGoogle Scholar
  12. 12.
    Wrighting DM, Andrews NC (2006) Interleukin-6 induces hepcidin expression through STAT3. Blood 108:3204–3209CrossRefPubMedGoogle Scholar
  13. 13.
    Pietrangelo A, Dierssen U, Valli L, Garuti C, Rump A, Corradini E, Ernst M, Klein C, Trautwein C (2007) STAT3 is required for IL-6-gp130-dependent activation of hepcidin in vivo. Gastroenterology 132:294–300CrossRefPubMedGoogle Scholar
  14. 14.
    Truksa J, Peng H, Lee P, Beutler E (2007) Different regulatory elements are required for response of hepcidin to interleukin-6 and bone morphogenetic proteins 4 and 9. Br J Haematol 139:138–147CrossRefPubMedGoogle Scholar
  15. 15.
    Truksa J, Lee P, Peng H, Flanagan J, Beutler E (2007) The distal location of the iron responsive region of the hepcidin promoter. Blood 110:3436–3437CrossRefPubMedGoogle Scholar
  16. 16.
    Xia Y, Babitt JL, Sidis Y, Chung RT, Lin HY (2008) Hemojuvelin regulates hepcidin expression via a selective subset of BMP ligands and receptors independently of neogenin. Blood 111:5195–5204CrossRefPubMedGoogle Scholar
  17. 17.
    Kautz L, Meynard D, Monnier A, Darnaud V, Bouvet R, Wang RH, Deng C, Vaulont S, Mosser J, Coppin H, Roth MP (2008) Iron regulates phosphorylation of Smad1/5/8 and gene expression of Bmp6, Smad7, Id1, and Atoh8 in the mouse liver. Blood 112:1503–1509CrossRefPubMedGoogle Scholar
  18. 18.
    Truksa J, Peng H, Lee P, Beutler E (2006) Bone morphogenetic proteins 2, 4, and 9 stimulate murine hepcidin 1 expression independently of Hfe, transferrin receptor 2 (Tfr2), and IL-6. Proc Natl Acad Sci U S A 103:10289–10293CrossRefPubMedGoogle Scholar
  19. 19.
    Cartharius K, Frech K, Grote K, Klocke B, Haltmeier M, Klingenhoff A, Frisch M, Bayerlein M, Werner T (2005) MatInspector and beyond: promoter analysis based on transcription factor binding sites. Bioinformatics 21:2933-2942CrossRefPubMedGoogle Scholar
  20. 20.
    Truksa J, Lee P, Beutler E (2008) Two BMP responsive elements, STAT, and bZIP/HNF4/COUP motifs of the hepcidin promoter are critical for BMP, SMAD1, and HJV responsiveness. Blood 113:688–695CrossRefPubMedGoogle Scholar
  21. 21.
    Niederkofler V, Salie R, Arber S (2005) Hemojuvelin is essential for dietary iron sensing, and its mutation leads to severe iron overload. J Clin Invest 115:2180–2186CrossRefPubMedGoogle Scholar
  22. 22.
    Roy CN, Custodio AO, de Graaf J, Schneider S, Akpan I, Montross LK, Sanchez M, Gaudino A, Hentze MW, Andrews NC, Muckenthaler MU (2004) An Hfe-dependent pathway mediates hyposideremia in response to lipopolysaccharide-induced inflammation in mice. Nat Genet 36:481–485CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Guillem Casanovas
    • 1
    • 1
    • 2
  • Katarzyna Mleczko-Sanecka
    • 1
    • 2
    • 3
  • Sandro Altamura
    • 1
    • 2
    • 3
  • Matthias W. Hentze
    • 2
    • 3
  • Martina U. Muckenthaler
    • 1
    • 2
  1. 1.Department of Pediatric Oncology, Hematology and ImmunologyUniversity of HeidelbergHeidelbergGermany
  2. 2.Molecular Medicine Partnership UnitEuropean Molecular Biology LaboratoryHeidelbergGermany
  3. 3.European Molecular Biology LaboratoryHeidelbergGermany

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