Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Dominant-negative mutations of CEBPA, encoding CCAAT/enhancer binding protein-α (C/EBPα), in acute myeloid leukemia

Nature Genetics volume 27, pages 263–270 (2001)Cite this article

Abstract

The transcription factor C/EBPα (for CCAAT/enhancer binding protein-α; encoded by the gene CEBPA) is crucial for the differentiation of granulocytes. Conditional expression of C/EBPα triggers neutrophilic differentiation, and no mature granulocytes are observed in Cebpa-mutant mice. Here we identify heterozygous mutations in CEBPA in ten patients with acute myeloid leukemia (AML). We found that five mutations in the amino terminus truncate the full-length protein, but did not affect a 30-kD protein initiated further downstream. The mutant proteins block wild-type C/EBPα DNA binding and transactivation of granulocyte target genes in a dominant-negative manner, and fails to induce granulocytic differentiation. Ours is the first report of CEBPA mutations in human neoplasia, and such mutations are likely to induce the differentiation block found in AML.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Diagrammatic representation of proteins encoded by wild-type and mutant CEBPA alleles.
Figure 2: Identification of proteins encoded by wild-type and mutated CEBPA alleles in vitro and in vivo.
Figure 3: DNA binding of C/EBPα mutant proteins and inhibition of wild-type C/EBPα binding.
Figure 4: Wild-type and mutant C/EBPα proteins are localized to the nucleus.
Figure 5: Transactivation potential of mutated CEBPA versus wild type.
Figure 6: Conditional expression of CEBPA mutant 263–269del fails to induce granulocytic differentiation and expression of CEBPA target genes in myeloid precursor cells.

Similar content being viewed by others

References

  1. Rabbits, T.H. Chromosomal translocations in human cancer. Nature 372, 143–149 (1994).

    Article  Google Scholar 

  2. Wolff, L. Contribution of oncogenes and tumor suppressor genes to myeloid leukemia. Biochim. Biophys. Acta 1332, F67–104 (1997).

    CAS  PubMed  Google Scholar 

  3. Tenen, D.G., Hromas, R., Licht, J.D. & Zhang, D.E. Transcription factors, normal myeloid development, and leukemia. Blood 90, 489–519 (1997).

    CAS  Google Scholar 

  4. Landschulz, W.H., Johnson, P.F., Adashi, E.Y., Graves, B.J. & McKnight, S.L. Isolation of a recombinant copy of the gene encoding C/EBP. Genes Dev. 2, 786–800 (1988).

    Article  CAS  Google Scholar 

  5. Cao, Z., Umek, R.M. & McKnight, S.L. Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells. Genes Dev. 5, 1538–1552 (1991).

    Article  CAS  Google Scholar 

  6. Lin, F.T. & Lane, M.D. CCAAT/enhancer binding protein α is sufficient to initiate the 3T3-L1 adipocyte differentiation program. Proc. Natl. Acad. Sci. USA 91, 8757–8761 (1994).

    Article  CAS  Google Scholar 

  7. Timchenko, N.A., Wilde, M., Nakanishi, M., Smith, J.R. & Darlington, G.J. CCAAT/enhancer-binding protein α (C/EBPα) inhibits cell proliferation through the p21 (WAF-1/CIP-1/SDI-1) protein. Genes Dev. 10, 804–815 (1996).

    Article  CAS  Google Scholar 

  8. Watkins, P.J., Condreay, J.P., Huber, B.E., Jacobs, S.J. & Adams, D.J. Impaired proliferation and tumorigenicity induced by CCAAT/enhancer-binding protein. Cancer Res. 56, 1063–1067 (1996).

    CAS  PubMed  Google Scholar 

  9. Umek, R.M., Friedman, A.D. & McKnight, S.L. CCAAT/enhancer binding protein α: a component of a differentiation switch. Science 251, 288–292 (1991).

    Article  CAS  Google Scholar 

  10. Hendricks-Taylor, L.R. & Darlington, G.J. Inhibition of cell proliferation by C/EBPα occurs in many cell types, does not require the presence of p53 or Rb, and is not affected by large T-antigen. Nucleic Acids Res. 23, 4726–4733 (1995).

    Article  CAS  Google Scholar 

  11. Scott, L.M., Civin, C.I., Roth, P. & Friedman, A.D. A novel temporal expression pattern of three C/EBP family members in differentiating myelomonocytic cells. Blood 80, 1725–1735 (1992).

    CAS  PubMed  Google Scholar 

  12. Radomska, H.S. et al. CCAAT/enhancer binding protein α is a regulatory switch sufficient for induction of granulocytic development from bipotential myeloid progenitors. Mol. Cell. Biol. 18, 4301–4314 (1998).

    Article  CAS  Google Scholar 

  13. Zhang, D.E. et al. Absence of granulocyte colony-stimulating factor signaling and neutrophil development in CCAAT enhancer binding protein α-deficient mice. Proc. Natl. Acad. Sci. USA 94, 569–574 (1997).

    Article  CAS  Google Scholar 

  14. An, M.R. et al. Evidence for posttranscriptional regulation of C/EBPα and C/EBPβ isoform expression during the lipopolysaccharide-mediated acute-phase response. Mol. Cell. Biol. 16, 2295–2306 (1996).

    Article  CAS  Google Scholar 

  15. Calkhoven, C.F., Bouwman, P.R.J., Snippe, L. & Ab, G. Translation start site multiplicity of the CCAAT/enhancer binding protein α mRNA is dictated by a small 5′ open reading frame. Nucleic Acids Res. 22, 5540–5547 (1994).

    Article  CAS  Google Scholar 

  16. Ossipow, V., Descombes, P. & Schibler, U. CCAAT/enhancer-binding protein mRNA is translated into multiple proteins with different transcription activation potentials. Proc. Natl. Acad. Sci. USA 90, 8219–8223 (1993).

    Article  CAS  Google Scholar 

  17. Smith, L.T., Hohaus, S., Gonzalez, D.A., Dziennis, S.E. & Tenen, D.G. PU.1 (Spi-1) and C/EBPα regulate the granulocyte colony-stimulating factor receptor promoter in myeloid cells. Blood 88, 1234–1247 (1996).

    CAS  PubMed  Google Scholar 

  18. Koike, M. et al. C/EBP-ɛ: chromosomal mapping and mutational analysis of the gene in leukemia and preleukemia. Leuk. Res. 21, 833–839 (1997).

    Article  CAS  Google Scholar 

  19. Yamanaka, R. et al. Impaired granulopoiesis, myelodysplasia, and early lethality in CCAAT/enhancer binding protein ɛ-deficient mice. Proc. Natl. Acad. Sci. USA 94, 13187–13192 (1997).

    Article  CAS  Google Scholar 

  20. Lekstrom-Himes, J.A., Dorman, S.E., Kopar, P., Holland, S.M. & Gallin, J.I. Neutrophil-specific granule deficiency results from a novel mutation with loss of function of the transcription factor CCAAT/enhancer binding protein ɛ. J. Exp. Med. 189, 1847–1852 (1999).

    Article  CAS  Google Scholar 

  21. Lekstrom-Himes, J.A. & Xanthopoulos, K.G. CCAAT/enhancer binding protein ɛ is critical for effective neutrophil-mediated response to inflammatory challenge. Blood 93, 3096–3105 (1999).

    CAS  PubMed  Google Scholar 

  22. Osato, M. et al. Biallelic and heterozygous point mutations in the Runt domain of the AML1/PEBP2αB gene associated with myeloblastic leukemias. Blood 93, 1817–1824 (1998).

    Google Scholar 

  23. Kozak, M. The scanning model for translation: an update. J. Cell Biol. 108, 229–241 (1989).

    Article  CAS  Google Scholar 

  24. Descombes, P. & Schibler, U. A liver-enriched transcriptional activator protein, LAP, and a transcriptional inhibitory protein, LIP, are translated from the same mRNA. Cell 67, 569–579 (1991).

    Article  CAS  Google Scholar 

  25. Lin, F.T., MacDougald, O.A., Diehl, A.M. & Lane, M.D. A 30-kD alternative translation product of the CCAAT/enhancer binding protein α message: transcriptional activator lacking antimitotic activity. Proc. Natl. Acad. Sci. USA 90, 9606–9610 (1993).

    Article  CAS  Google Scholar 

  26. Hendricks-Taylor, L.R. & Darlington, G.J. The CCAAT/enhancer-binding protein (C/EBP α) gene (CEBPA) maps to human chromosome 19q13.1 and the related nuclear factor NF-IL6 (C/EBP β) gene (CEBPB) gene maps to human chromosome 20q13.1. Genomics 14, 12–17 (1992).

    Article  CAS  Google Scholar 

  27. Westendorf, J.J. et al. The t(8;21) fusion product, AML-1-ETO, associates with C/EBPα, inhibits C/EBPα-dependent transcription, and blocks granulocytic differentiation. Mol. Cell. Biol. 18, 322–333 (1998).

    Article  CAS  Google Scholar 

  28. Swart, G.W.M., van Groningen, J.J.M., van Ruissen, F., Bergers, M. & Schalkwijk, J. Transcription factor C/EBPα: novel sites of expression and cloning of the human gene. Biol. Chem. 378, 373–379 (1997).

    Article  CAS  Google Scholar 

  29. Pahl, H.L., Burn, T.C. & Tenen, D.G. Optimization of transient transfection into human myeloid cell lines using a luciferase reporter gene. Exp. Hematol. 19, 1038–1041 (1991).

    CAS  PubMed  Google Scholar 

  30. Nerlov, C. & Ziff, E.B. Three levels of functional interaction determine the activity of CCAAT/enhancer binding protein-α on the serum albumin promoter. Genes Dev. 8, 350–362 (1994).

    Article  CAS  Google Scholar 

  31. Friedman, A.D. & McKnight, S.L. Identification of two polypeptide segments of CCAAT/enhancer-binding protein required for transcriptional activation of the serum albumin gene. Genes Dev. 4, 1416–1426 (1990).

    Article  CAS  Google Scholar 

  32. Landschulz, W.H., Johnson, P.F. & McKnight, S.L. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science 240, 1759–1764 (1988).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank C. Schoch for assistance with cytogenetic analysis and T. Haferlach for morphologic analysis of patient samples; members of the Tenen laboratory, A. Iwama and G. Gilliland for suggestions; L. Clayton for careful reading of the manuscript; and M. Singleton for assistance with preparation of the manuscript. This work was supported by grants from the Swiss National Science Foundation (81BS-051911 and SSMBS 1011) and the Swiss Roche Research Foundation (97-189) (to T.P.), and NIH grants CA72009 and HL56745 (to D.G.T.).

Author information

Authors and Affiliations

  1. Hematology/Oncology Division, Harvard Institutes of Medicine, Harvard Medical School, Boston, Massachusetts, USA

    Thomas Pabst, Beatrice U. Mueller, Pu Zhang, Hanna S. Radomska, Sailaja Narravula & Daniel G. Tenen

  2. Department of Medicine III, Grosshadern, Ludwig Maximilians University Munich, Munich, Germany

    Susanne Schnittger, Gerhard Behre & Wolfgang Hiddemann

Authors
  1. Thomas Pabst
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Beatrice U. Mueller
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pu Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hanna S. Radomska
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sailaja Narravula
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Susanne Schnittger
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gerhard Behre
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Wolfgang Hiddemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Daniel G. Tenen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Daniel G. Tenen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pabst, T., Mueller, B., Zhang, P. et al. Dominant-negative mutations of CEBPA, encoding CCAAT/enhancer binding protein-α (C/EBPα), in acute myeloid leukemia. Nat Genet 27, 263–270 (2001). https://doi.org/10.1038/85820

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/85820

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing