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Positional cloning of the zebrafish sauternes gene: a model for congenital sideroblastic anaemia

Nature Genetics volume 20, pages 244–250 (1998)Cite this article

Abstract

Many human anaemias are caused by defects in haemoglobin synthesis. The zebrafish mutant sauternes (sau) has a microcytic, hypochromic anaemia, suggesting that haemoglobin production is perturbed. During embryogenesis, sau mutants have delayed erythroid maturation and abnormal globin gene expression. Using positional cloning techniques, we show that sau encodes the erythroid-specific isoform of δ-aminolevulinate synthase (ALAS2; also known as ALAS-E), the enzyme required for the first step in haem biosynthesis. As mutations in ALAS2 cause congenital sideroblastic anaemia (CSA) in humans, sau represents the first animal model of this disease.

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Figure 1: Wright-Giemsa staining of circulating embryonic red blood cells from wild-type and sau animals.
Figure 2: Analysis of haemoglobin, embryonic globin and gata1 expression in sau mutant embryos.
Figure 3: Haematologic phenotype of sau adults.
Figure 4: Haem content of sauty121 red blood cells.
Figure 5: alas2 is the sau gene.
Figure 6: Expression of wild-type alas2 cDNA partially rescued the sau phenotype.
Figure 7: Expression analysis of alas1 and alas2 in the zebrafish embryo.

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Acknowledgements

We thank A. Deconinck, S. Ganiatsas and P. Mead for critical review of this manuscript; V. Deane, L. Garbaczewski, C. Rogers and B. Vail for technical assistance. We thank P. Haffter and C. Nusslein-Volhard for providing zebrafish blood mutants before publication, C. Amemiya and G. Silverman for assistance in working with PAC and YAC clones, respectively, and M. Fleming for assistance with prussian blue staining. M. Clark and H. Lerach generously provided kidney cDNA library filters. We also thank Z. Gong for providing the alas1 EST. C. Shackleton provided support for globin structural studies using a VG BioQ mass spectrometer (RR06505). We thank L. Kunkel, S. Orkin and W. Talbot for their support and advice. This work was funded by NIH grants 1RO1 DK53298-01 and P50 DK49216-05 and by USPHS grant DK-32890. H.E.W. is supported through the Northern California Comprehensive Sickle Cell Center Grant No. HL20985. B.H.P. is supported by a Howard Hughes Postdoctoral Fellowship for Physicians. A.C.O. is the recipient of an Anti-Cancer Victoria Postgraduate Research Scholarship. L.I.Z. is an Associate Investigator of the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Division of Hematology/Oncology, Department of Pediatrics, Children's Hospital, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Alison Brownlie, Adriana Donovan, Stephen J. Pratt, Barry H. Paw, Carlo Brugnara & Leonard I. Zon

  2. Howard Hughes Medical Institute, Boston, 02115, Massachusetts, USA

    Barry H. Paw & Leonard I. Zon

  3. Ludwig Institute for Cancer Research, PO Royal Melbourne Hospital, VIC, Australia

    Andrew C. Oates

  4. Children's Hospital Oakland Research Institute, Oakland, 94609, California, USA

    H. Ewa Witkowska

  5. The Rockefeller University, New York, 10021, New York, USA

    Shigeru Sassa

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Brownlie, A., Donovan, A., Pratt, S. et al. Positional cloning of the zebrafish sauternes gene: a model for congenital sideroblastic anaemia. Nat Genet 20, 244–250 (1998). https://doi.org/10.1038/3049

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