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Abstract

Iron has a fundamental role in many metabolic processes, including electron transport, deoxyribonucleotide synthesis, oxygen transport and many essential redox reactions involving haemoproteins and Fe–S cluster proteins. Defective iron homeostasis results in either iron deficiency or iron overload1. Precise regulation of iron transport in mitochondria is essential for haem biosynthesis2, haemoglobin production and Fe–S cluster protein assembly3,4 during red cell development. Here we describe a zebrafish mutant, frascati (frs)5, that shows profound hypochromic anaemia and erythroid maturation arrest owing to defects in mitochondrial iron uptake. Through positional cloning, we show that the gene mutated in the frs mutant is a member of the vertebrate mitochondrial solute carrier family (SLC25)6 that we call mitoferrin (mfrn). mfrn is highly expressed in fetal and adult haematopoietic tissues of zebrafish and mouse. Erythroblasts generated from murine embryonic stem cells null for Mfrn (also known as Slc25a37) show maturation arrest with severely impaired incorporation of 55Fe into haem. Disruption of the yeast mfrn orthologues, MRS3 and MRS4, causes defects in iron metabolism and mitochondrial Fe–S cluster biogenesis7,8,9,10. Murine Mfrn rescues the defects in frs zebrafish, and zebrafish mfrn complements the yeast mutant, indicating that the function of the gene may be highly conserved. Our data show that mfrn functions as the principal mitochondrial iron importer essential for haem biosynthesis in vertebrate erythroblasts.

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Acknowledgements

We thank C. Lawrence, A. Walker and C. Belair for zebrafish animal husbandry; M. Halpern for sptb333 zebrafish; S. Johnson for SJD zebrafish; W. Shu Wu for mouse bone marrow cDNA; C. Roy for advice on preparing 55Fe-saturated transferrin; M. Kaku and Y. Fujiwara for advice on ES cell culture and selection; H. Wohlrab for advice on SLC25 biochemistry; K. Dooley for the frsij001 allele from the Tübingen 2000 screen; M. Ocaña for help with confocal fluorescence images; S. Dallaire and C. Lee for karyotyping the Mfrn-null ES cells; A. B. Cantor for advice on generating Hox-11-immortalized haematopoietic cells; and E. Shafizadeh, S.-K. Choe, C. Burns, Y. Houvras, D. Langenau, W. Tse, H. Wolhrab, J. Kanki and G. M. Shaw for critically reading the manuscript. This work was supported in part by the William Randolph Hearst Foundation (B.H.P.); the March of Dimes Birth Defects Foundation Basil O'Connor Award (B.H.P.); the Swiss National Science Foundation (G.E.A.); the Belgian National Research Fund (E.M.); the NIH (L.L.P., L.I.Z., J.K., M.J.W., B.H.P.); and the Howard Hughes Medical Institute (L.I.Z.).

Author information

Author notes

    • George C. Shaw
    • , John J. Cope
    • , Gabriele E. Ackermann
    • , Rebecca A. Wingert
    • , David Traver
    • , Nikolaus S. Trede
    •  & Alison Brownlie

    †Present addresses: Ohio State University College of Medicine, Columbus, Ohio 43210, USA (G.C.S.); University at Buffalo School of Medicine and Biomedical Sciences, Buffalo, New York 14214, USA (J.J.C.); Kinderspital Zürich, Zürich 8032, Switzerland (G.E.A.); Massachusetts General Hospital, Boston, Massachusetts 02114, USA (R.A.W.); University of California, San Diego, California 92093, USA. (D.T.); University of Utah, Salt Lake City, Utah 84112, USA (N.S.T.); Xenon Pharmaceuticals, Burnaby, British Columbia V5G 4W8, Canada (A.B.)

Affiliations

  1. Division of Hematology, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA

    • George C. Shaw
    • , John J. Cope
    • , Kenneth Corson
    • , Gabriele E. Ackermann
    • , Emmanuel Minet
    •  & Barry H. Paw
  2. Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah 84132, USA

    • Liangtao Li
    •  & Jerry Kaplan
  3. Howard Hughes Medical Institute, Stem Cell Program and Division of Hematology-Oncology, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Candace Hersey
    • , Rebecca A. Wingert
    • , David Traver
    • , Nikolaus S. Trede
    • , Bruce A. Barut
    • , Yi Zhou
    • , Adriana Donovan
    • , Alison Brownlie
    •  & Leonard I. Zon
  4. The Jackson Laboratory, Bar Harbor, Maine 04609, USA

    • Babette Gwynn
    • , Amy J. Lambert
    •  & Luanne L. Peters
  5. Department of Physiology and Biochemistry, University of Malta, Msida MSD 06, Malta

    • Rena Balzan
  6. Division of Hematology, Department of Pediatrics, Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA

    • Mitchell J. Weiss

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Competing interests

Sequences have been deposited in GenBank as follows: Zebrafish mfrn (slc25a37; DQ112224, DQ112225), Zebrafish mfrn2 (slc25a28; BC054641), mouse Mfrn (Slc25a37; AF361699), mouse Mfrn2 (Slc25a28; BC025908.1), human MFRN (MSCP; SLC25A37; NM_016612) and human MFRN2 (SLC25A28; BC058937). Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Corresponding author

Correspondence to Barry H. Paw.

Supplementary information

PDF files

  1. 1.

    Supplementary Figure 1.

    Analysis of the frs anemic phenotype and the expression of mfrn in zebrafish

  2. 2.

    Supplementary Figure 2.

    Expression pattern of murine Mfrn in cultured cells and zebrafish mfrn2 in developing embryos.

  3. 3.

    Supplementary Figure 3.

    Biochemical characterization of mfrn-deficient mouse hematopoietic cells and yeasts.

  4. 4.

    Supplementary Figure 4.

    Zebrafish mfrn corrects the mitochondrial iron deficiency of yeast Δmrs3/4 mutant.

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    Supplementary Figure Legends

    Text to accompany the above Supplementary Figures.

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DOI

https://doi.org/10.1038/nature04512

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