A transcription-factor-binding surface of coactivator p300 is required for haematopoiesis

Abstract

The coactivators CBP (Cre-element binding protein (CREB)-binding protein) and its paralogue p300 are thought to supply adaptor molecule and protein acetyltransferase functions to many transcription factors that regulate gene expression1. Normal development requires CBP and p300, and mutations in these genes are found in haematopoietic and epithelial tumours2,3,4,5,6. It is unclear, however, which functions of CBP and p300 are essential in vivo. Here we show that the protein-binding KIX domains of CBP and p300 have nonredundant functions in mice. In mice homozygous for point mutations in the KIX domain of p300 designed to disrupt the binding surface for the transcription factors c-Myb and CREB7,8,9, multilineage defects occur in haematopoiesis, including anaemia, B-cell deficiency, thymic hypoplasia, megakaryocytosis and thrombocytosis. By contrast, age-matched mice homozygous for identical mutations in the KIX domain of CBP are essentially normal. There is a synergistic genetic interaction between mutations in c-Myb and mutations in the KIX domain of p300, which suggests that the binding of c-Myb to this domain of p300 is crucial for the development and function of megakaryocytes. Thus, conserved domains in two highly related coactivators have contrasting roles in haematopoiesis.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Mouse KIX domain mutations.
Figure 2: Haematopoietic defects in p300KIX/KIX mice.
Figure 3: Mutant CBP and p300 affect CREB and c-Myb.
Figure 4: c-Myb and p300 KIX functionally interact to regulate megakaryocytopoiesis.

References

  1. 1

    Vo, N. K. & Goodman, R. H. CREB-binding protein and p300 in transcriptional regulation. J. Biol. Chem. 276, 13505–13508 (2001)

    CAS  Article  Google Scholar 

  2. 2

    Oike, Y. et al. Mice homozygous for a truncated form of CREB-binding protein exhibit defects in hematopoiesis and vasculo-angiogenesis. Blood 93, 2771–2779 (1999)

    CAS  PubMed  Google Scholar 

  3. 3

    Yao, T. P. et al. Gene dosage-dependent embryonic development and proliferation defects in mice lacking the transcriptional integrator p300. Cell 93, 361–372 (1998)

    CAS  Article  Google Scholar 

  4. 4

    Kung, A. L. et al. Gene dose-dependent control of hematopoiesis and hematologic tumor suppression by CBP. Genes Dev. 14, 272–277 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  5. 5

    Gayther, S. A. et al. Mutations truncating the EP300 acetylase in human cancers. Nature Genet. 24, 300–303 (2000)

    CAS  Article  Google Scholar 

  6. 6

    Goodman, R. H. & Smolik, S. CBP/p300 in cell growth, transformation, and development. Genes Dev. 14, 1553–1577 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7

    Parker, D. et al. Analysis of an activator:coactivator complex reveals an essential role for secondary structure in transcriptional activation. Mol. Cell 2, 353–359 (1998)

    CAS  Article  Google Scholar 

  8. 8

    Parker, D. et al. Role of secondary structure in discrimination between constitutive and inducible activators. Mol. Cell. Biol. 19, 5601–5607 (1999)

    CAS  Article  Google Scholar 

  9. 9

    Radhakrishnan, I. et al. Solution structure of the KIX domain of CBP bound to the transactivation domain of CREB: a model for activator:coactivator interactions. Cell 91, 741–752 (1997)

    CAS  Article  Google Scholar 

  10. 10

    Jones, D. T. Protein secondary structure prediction based on position-specific scoring matrices. J. Mol. Biol. 292, 195–202 (1999)

    CAS  Article  Google Scholar 

  11. 11

    Munoz, V. & Serrano, L. Elucidating the folding problem of helical peptides using empirical parameters. II. Helix macrodipole effects and rational modification of the helical content of natural peptides. J. Mol. Biol. 245, 275–296 (1995)

    CAS  Article  Google Scholar 

  12. 12

    Kraus, W. L., Manning, E. T. & Kadonaga, J. T. Biochemical analysis of distinct activation functions in p300 that enhance transcription initiation with chromatin templates. Mol. Cell. Biol. 19, 8123–8135 (1999)

    CAS  Article  Google Scholar 

  13. 13

    Yang, C., Shapiro, L. H., Rivera, M., Kumar, A. & Brindle, P. K. A role for CREB binding protein and p300 transcriptional coactivators in Ets-1 transactivation functions. Mol. Cell. Biol. 18, 2218–2229 (1998)

    CAS  Article  Google Scholar 

  14. 14

    Jayaraman, G. et al. p300/cAMP-responsive element-binding protein interactions with ets-1 and ets-2 in the transcriptional activation of the human stromelysin promoter. J. Biol. Chem. 274, 17342–17352 (1999)

    CAS  Article  Google Scholar 

  15. 15

    Mucenski, M. L. et al. A functional c-myb gene is required for normal murine fetal hepatic hematopoiesis. Cell 65, 677–689 (1991)

    CAS  Article  Google Scholar 

  16. 16

    Allen, R. D., Bender, T. P. & Siu, G. c-Myb is essential for early T cell development. Genes Dev. 13, 1073–1078 (1999)

    CAS  Article  Google Scholar 

  17. 17

    Herschlag, D. & Johnson, F. B. Synergism in transcriptional activation: a kinetic view. Genes Dev. 7, 173–179 (1993)

    CAS  Article  Google Scholar 

  18. 18

    Hartman, J. L., Garvik, B. & Hartwell, L. Principles for the buffering of genetic variation. Science 291, 1001–1004 (2001)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Jacobsson, S. et al. Flow cytometric analysis of megakaryocyte ploidy in chronic myeloproliferative disorders and reactive thrombocytosis. Eur. J. Haematol. 56, 287–292 (1996)

    CAS  Article  Google Scholar 

  20. 20

    Jackson, C. W., Steward, S. A., Chenaille, P. J., Ashmun, R. A. & McDonald, T. P. An analysis of megakaryocytopoiesis in the C3H mouse: an animal model whose megakaryocytes have 32N as the modal DNA class. Blood 76, 690–696 (1990)

    CAS  PubMed  Google Scholar 

  21. 21

    Sumner, R., Crawford, A., Mucenski, M. & Frampton, J. Initiation of adult myelopoiesis can occur in the absence of c-Myb whereas subsequent development is strictly dependent on the transcription factor. Oncogene 19, 3335–3342 (2000)

    CAS  Article  Google Scholar 

  22. 22

    Rudolph, D. et al. Impaired fetal T cell development and perinatal lethality in mice lacking the cAMP response element binding protein. Proc. Natl Acad. Sci. USA 95, 4481–4486 (1998)

    ADS  CAS  Article  Google Scholar 

  23. 23

    Arnold, J. T. et al. A single injection of pegylated murine megakaryocyte growth and development factor (MGDF) into mice is sufficient to produce a profound stimulation of megakaryocyte frequency, size, and ploidization. Blood 89, 823–833 (1997)

    CAS  PubMed  Google Scholar 

  24. 24

    Kasper, L. H. et al. CREB binding protein interacts with nucleoporin-specific FG repeats that activate transcription and mediate NUP98-HOXA9 oncogenicity. Mol. Cell. Biol. 19, 764–776 (1999)

    CAS  Article  Google Scholar 

  25. 25

    Shapiro, L. H. Myb and Ets proteins cooperate to transactivate an early myeloid gene. J. Biol. Chem. 270, 8763–8771 (1995)

    CAS  Article  Google Scholar 

  26. 26

    Eckner, R. et al. Molecular cloning and functional analysis of the adenovirus E1A-associated 300-kD protein (p300) reveals a protein with properties of a transcriptional adaptor. Genes Dev. 8, 869–884 (1994)

    CAS  Article  Google Scholar 

  27. 27

    Lecine, P., Blank, V. & Shivdasani, R. Characterization of the hematopoietic transcription factor NF-E2 in primary murine megakaryocytes. J. Biol. Chem. 273, 7572–7578 (1998)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We thank M. Castor, D. Bush, S. Kelly and M. Harper for technical help; J. Gatewood, M. Paktinat, R. Cross, R. Ashmun and A. Hoffmeyer for FACS analyses; S. Steward and T. Pestina for help with the platelet counts; R. Piekorz for assistance with the bone marrow transplants; X. Xiong and J. Schroeder for advice on statistics; A. Hoffmeyer for discussions; R. Shivdasani for advice on purifying megakaryocytes; H. Singh for c-Myb mice; and the Transgenic/Gene Knockout Shared Resource for technical assistance. This work was supported by a grant from the NIH and from the National Cancer Institute Cancer Center Support (CORE) program, and by the American Lebanese Syrian Associated Charities of St Jude Children's Research Hospital.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Paul K. Brindle.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kasper, L., Boussouar, F., Ney, P. et al. A transcription-factor-binding surface of coactivator p300 is required for haematopoiesis. Nature 419, 738–743 (2002). https://doi.org/10.1038/nature01062

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Sign up for the Nature Briefing newsletter for a daily update on COVID-19 science.
Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing