An inherited mutation leading to production of only the short isoform of GATA-1 is associated with impaired erythropoiesis (original) (raw)

Nature Genetics volume 38, pages 807–812 (2006)Cite this article

Abstract

Acquired somatic mutations1,2,3,4,5,6 in exon 2 of the hematopoietic transcription factor GATA-1 have been found in individuals with Down syndrome with both transient myeloproliferative disorder2,3,4,5,6,7,8,9,10 and acute megakaryoblastic leukemia1,2,3,4,6,8,9,10. These mutations prevent the synthesis of the full-length protein but allow the synthesis of its short isoform, GATA-1s. Experiments in mice11 suggest that GATA-1s supports normal adult megakaryopoiesis, platelet formation and erythropoiesis. Here we report a mutation, 332G → C, in exon 2 of GATA1, leading to the synthesis of only the short isoform in seven affected males from two generations of a family. Hematological profiles of affected males demonstrate macrocytic anemia, normal platelet counts and neutropenia in most cases. Altogether, data suggest that GATA-1s alone, produced in low or normal levels, is not sufficient to support normal erythropoiesis. Moreover, this is the first study to indicate that a germline splicing mutation does not lead to leukemia in the absence of other cooperating events, such as Down syndrome.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 12 print issues and online access

$209.00 per year

only $17.42 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Wechsler, J. et al. Acquired mutations in GATA1 in the megakaryoblastic leukemia of Down syndrome. Nat. Genet. 32, 148–152 (2002).
    Article CAS Google Scholar
  2. Hitzler, J.K., Cheung, J., Li, Y., Scherer, S.W. & Zipursky, A. GATA1 mutations in transient leukemia and acute megakaryoblastic leukemia of Down syndrome. Blood 101, 4301–4304 (2003).
    Article CAS Google Scholar
  3. Rainis, L. et al. Mutations in exon 2 of GATA1 are early events in megakaryocytic malignancies associated with trisomy 21. Blood 102, 981–986 (2003).
    Article CAS Google Scholar
  4. Mundschau, G. et al. Mutagenesis of GATA1 is an initiating event in Down syndrome leukemogenesis. Blood 101, 4298–4300 (2003).
    Article CAS Google Scholar
  5. Xu, G. et al. Frequent mutations in the GATA-1 gene in the transient myeloproliferative disorder of Down syndrome. Blood 102, 2960–2968 (2003).
    Article CAS Google Scholar
  6. Ahmed, M. et al. Natural history of GATA1 mutations in Down syndrome. Blood 103, 2480–2489 (2004).
    Article CAS Google Scholar
  7. Groet, J. et al. Acquired mutations in GATA1 in neonates with Down's syndrome with transient myeloid disorder. Lancet 361, 1617–1620 (2003).
    Article CAS Google Scholar
  8. Hitzler, J.K. & Zipursky, A. A. Origins of leukaemia in children with Down syndrome. Nat. Rev. Cancer 5, 11–20 (2005).
    Article CAS Google Scholar
  9. Shimada, A. et al. Fetal origin of the GATA1 mutation in identical twins with transient myeloproliferative disorder and acute megakaryoblastic leukemia accompanying Down syndrome. Blood 103, 366 (2004).
    Article CAS Google Scholar
  10. Crispino, J.D. GATA1 mutations in Down syndrome: implications for biology and diagnosis of children with transient myeloproliferative disorder and acute megakaryoblastic leukemia. Pediatr. Blood Cancer 44, 40–44 (2005).
    Article Google Scholar
  11. Li, Z. et al. Developmental stage-selective effect of somatically mutated leukemogenic transcription factor GATA1. Nat. Genet. 37, 613–619 (2005).
    Article CAS Google Scholar
  12. Nichols, K.E. et al. Familial dyserythropoietic anaemia and thrombocytopenia due to an inherited mutation in GATA1 . Nat. Genet. 24, 266–270 (2000).
    Article CAS Google Scholar
  13. Mehaffey, M.G., Newton, A.L., Gandhi, M.J., Crossley, M. & Drachman, J.G. X-linked thrombocytopenia caused by a novel mutation of GATA-1. Blood 98, 2681–2688 (2001).
    Article CAS Google Scholar
  14. Freson, K. et al. Platelet characteristics in patients with X-linked macrothrombocytopenia because of a novel GATA1 mutation. Blood 98, 85–92 (2001).
    Article CAS Google Scholar
  15. Yu, C. et al. X-linked thrombocytopenia with thalassemia from a mutation in the amino finger of GATA-1 affecting DNA binding rather than FOG-1 interaction. Blood 100, 2040–2045 (2002).
    Article CAS Google Scholar
  16. Balduini, C.L. et al. Effects of the R216Q mutation of GATA-1 on erythropoiesis and megakaryocytopoiesis. Thromb. Haemost. 91, 129–140 (2004).
    Article CAS Google Scholar
  17. Freson, K. et al. Different substitutions at residue D218 of the X-linked transcription factor GATA1 lead to altered clinical severity of macrothrombocytopenia and anemia and are associated with variable skewed X inactivation. Hum. Mol. Genet. 11, 147–152 (2002).
    Article CAS Google Scholar
  18. Calligaris, R., Bottardi, S., Cogoi, S., Apezteguia, I. & Santoro, C. Alternative translation initiation site usage results in two functionally distinct forms of the GATA-1 transcription factor. Proc. Natl. Acad. Sci. USA 92, 11598–11602 (1995).
    Article CAS Google Scholar
  19. Gurbuxani, S., Vyas, P. & Crispino, J.D. Recent insights into the mechanisms of myeloid leukemogenesis in Down syndrome. Blood 103, 399–406 (2004).
    Article CAS Google Scholar
  20. Shimizu, R., Takahashi, S., Ohneda, K., Engel, J.D. & Yamamoto, M. In vivo requirements for GATA-1 functional domains during primitive and definitive erythropoiesis. EMBO J. 20, 5250–5260 (2001).
    Article CAS Google Scholar
  21. Rhodes, J. et al. Interplay of pu.1 and gata1 determines myelo-erythroid progenitor cell fate in zebrafish. Dev. Cell 8, 97–108 (2005).
    Article CAS Google Scholar
  22. Centurione, L. et al. Increased and pathological emperipolesis of neutrophils within megakaryocytes associated with marrow fibrosis in GATA-1low mice. Blood 104, 3573–3580 (2004).
    Article CAS Google Scholar
  23. Conran, N., Fattori, A., Saad, S.T. & Costa, F.F. Increased levels of soluble ICAM-1 in the plasma of sickle cell patients are reversed by hydroxyurea. Am. J. Hematol. 76, 343–347 (2004).
    Article CAS Google Scholar
  24. dos Santos, C.O., Duarte, A.S., Saad, S.T. & Costa, F.F. Expression of alpha-hemoglobin stabilizing protein gene during human erythropoiesis. Exp. Hematol. 32, 157–162 (2004).
    Article Google Scholar
  25. Vandesompele, J. et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3 Research0034.1–Research0034.11 (2002).
  26. Budde, U., Drewke, E., Mainusch, K. & Schneppenheim, R. Laboratory diagnosis of congenital von Willebrand disease. Semin. Thromb. Hemost. 28, 173–190 (2002).
    Article CAS Google Scholar
  27. Gawaz, M. & Ward, R. Effects of hemodialysis on platelet-derived thrombospondin. Kid. Inter. 40, 257–265 (1991).
    Article CAS Google Scholar
  28. Gao, D.Y. et al. Development of optimal techniques for cryopreservation of human platelets. I. Platelet activation during cold storage (at 22 and 8 degrees C) and cryopreservation. Cryobiology 38, 225–235 (1999).
    Article CAS Google Scholar
  29. Pedrazzoli, P. et al. Transfusion of platelet concentrates cryopreserved with thrombosol plus low-dose dimethylsulphoxide in patients with severe thrombocytopenia: a pilot study. Br. J. Haematol. 108, 653–659 (2000).
    Article CAS Google Scholar
  30. Stenberg, P.E. et al. Prolonged bleeding time with defective platelet filopodia formation in the Wistar Furth rat. Blood 91, 1599–1608 (1998).
    CAS PubMed Google Scholar

Download references

Acknowledgements

We thank N. Conran for her English revision of the manuscript and suggestions; S. Gambero and C. Lanaro for technical assistance with Genorm analysis and Real Time PCR; C.A. Chagas, J.C. Moraes, T.E. Prates for technical assistance with the immunohistochemical study in bone marrow biopsies; R. Secolin for technical assistance with Lod Score analyses; T. Machado for technical assistance with the analysis of plasma coagulation factors and the plasma coagulation assay; F.G. Pereira for technical assistance with flow cytometric analysis of platelet membrane glycoproteins in platelet-rich plasma and J.L. Saturno for technical assistance with ultrastructural analysis of platelets. This work was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP; grant no. 02/13801-7); Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; grant no. 301310/2004-1) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES; grant no. DS-108/00).

Author information

Authors and Affiliations

  1. Department of Internal Medicine, Hemocentro, School of Medical Science, Universidade Estadual de Campinas, Campinas, São Paulo, 13083-970, Brazil
    Luciana M Hollanda, Carmen S P Lima, Anderson F Cunha, Dulcinéia M Albuquerque, Margareth C Ozelo, Sara T O Saad & Fernando F Costa
  2. Department of Pathological Anatomy, School of Medical Sciences, Campinas, São Paulo, 13083-970, Brazil
    José Vassallo
  3. Department of Histology, Institute of Biology, Universidade Estadual de Campinas, Campinas, São Paulo, 13083-970, Brazil
    Paulo P Joazeiro

Authors

  1. Luciana M Hollanda
    You can also search for this author inPubMed Google Scholar
  2. Carmen S P Lima
    You can also search for this author inPubMed Google Scholar
  3. Anderson F Cunha
    You can also search for this author inPubMed Google Scholar
  4. Dulcinéia M Albuquerque
    You can also search for this author inPubMed Google Scholar
  5. José Vassallo
    You can also search for this author inPubMed Google Scholar
  6. Margareth C Ozelo
    You can also search for this author inPubMed Google Scholar
  7. Paulo P Joazeiro
    You can also search for this author inPubMed Google Scholar
  8. Sara T O Saad
    You can also search for this author inPubMed Google Scholar
  9. Fernando F Costa
    You can also search for this author inPubMed Google Scholar

Contributions

L.M.H. performed and processed most of experiments and wrote the first draft of the manuscript. C.S.P.L. contributed for the clinical and bone marrow morphology data of individuals. A.F.C. and D.M.A. performed some experiments and assisted L.M.H. in a number of procedures. J.V. contributed to the morphological and immunohistochemical study in bone marrow biopsies. M.C.O. contributed to the study of plasma coagulation factors, platelet aggregation assays and flow cytometric analysis of platelet membrane glycoproteins in platelet-rich plasma. P.P.J. contributed for the ultrastructural analysis of platelets. S.T.O.S. contributed to analysis of the clinical and laboratory data. F.F.C. conceived and designed the study and was responsible for the final draft of the manuscript. All authors were involved in the final revision of the article, contributed to the interpretation of the data and gave final approval to the manuscript.

Corresponding author

Correspondence toFernando F Costa.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Relative expression of mRNA obtained by real-time PCR of GATA1 and GATA1s from K562 cells, individual III-16, and pooled normal bone marrow cells. (PDF 83 kb)

Supplementary Table 1

Clinical laboratory findings and morphological characteristics of peripheral blood and bone marrow in affected males. (PDF 23 kb)

Supplementary Table 2

Hematological data from heterozygous families. (PDF 27 kb)

Supplementary Table 3

Platelet functional studies of the unaffected, heterozygous and hemizygous family members. (PDF 30 kb)

Supplementary Table 4

Primer sequences for PCR and RT-PCR. (PDF 12 kb)

Supplementary Table 5

Primer sequences for real-time quantitative PCR. (PDF 14 kb)

Rights and permissions

About this article

Cite this article

Hollanda, L., Lima, C., Cunha, A. et al. An inherited mutation leading to production of only the short isoform of GATA-1 is associated with impaired erythropoiesis.Nat Genet 38, 807–812 (2006). https://doi.org/10.1038/ng1825

Download citation

This article is cited by

Associated content