Iron is required to produce haem and iron–sulphur (Fe–S) clusters, processes thought to occur independently1,2. Here we show that the hypochromic anaemia in shiraz (sir) zebrafish mutants is caused by deficiency of glutaredoxin 5 (grx5), a gene required in yeast for Fe–S cluster assembly. We found that grx5 was expressed in erythroid cells of zebrafish and mice. Zebrafish grx5 rescued the assembly of Δgrx5 yeast Fe–S, showing that the biochemical function of grx5 is evolutionarily conserved. In contrast to yeast, vertebrates use iron regulatory protein 1 (IRP1) to sense intracellular iron and regulate mRNA stability or the translation of iron metabolism genes1,2. We found that loss of Fe–S cluster assembly in sir animals activated IRP1 and blocked haem biosynthesis catalysed by aminolaevulinate synthase 2 (ALAS2). Overexpression of ALAS2 RNA without the 5′ iron response element that binds IRP1 rescued sir embryos, whereas overexpression of ALAS2 including the iron response element did not. Further, antisense knockdown of IRP1 restored sir embryo haemoglobin synthesis. These findings uncover a connection between haem biosynthesis and Fe–S clusters, indicating that haemoglobin production in the differentiating red cell is regulated through Fe–S cluster assembly.
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We thank all members of the Zon laboratory for their advice and support. R.A.W. is a Harvard University Fellow of the Albert J. Ryan Foundation. L.I.Z. is an Investigator of the Howard Hughes Medical Institute. This work was supported by HHMI and NIH grants.
The zebrafish grx5, murine grx5, human grx5, zebrafish IRP1 and zebrafish IRP2 sequences are deposited in GenBank under accession numbers DQ083329, DQ083330, DQ083331, DQ083332 and DQ083333, respectively. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
A list of members of the Tübingen 2000 Screen Consortium and their affiliations appears at the end of the paper
The subcellular localization of zebrafish grx5 protein when zgrx5 was overexpressed in HEK293 cells.
This figure shows sir-/- zebrafish that were raised to juvenile stage when zebrafish grx5 mRNA was overexpressed during embryogenesis.
This figure shows the in situ hybridization staining for expression of zebrafish IRP1 and IRP2 mRNA at 24 hpf.
Legends to accompany the above Supplementary Figures.
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Wingert, R., Galloway, J., Barut, B. et al. Deficiency of glutaredoxin 5 reveals Fe–S clusters are required for vertebrate haem synthesis. Nature 436, 1035–1039 (2005). https://doi.org/10.1038/nature03887
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