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M408654200v1
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Papers In Press, published online ahead of print October 26, 2004
J. Biol. Chem, 10.1074/jbc.M408654200
Submitted on July 29, 2004
Revised on October 26, 2004
Accepted on October 26, 2004

E2A-PBX1 interacts directly with the KIX domain of CBP/p300 in the induction of proliferation in primary hematopoietic cells

R. Bayly, L. Chuen, R.A. Currie, B.D. Hyndman, R. Casselman, G.A. Blobel, and D.P. LeBrun

Pathology and Molecular Medicine, Queen's University, Kingston, Ontario K7L 3N6

Corresponding Author: lebrun{at}cliff.path.queensu.ca

The E2A gene encodes DNA-binding transcription factors, called E12 and E47, involved in cell specification and maturation. E2A is also involved in a chromosomal translocation that leads to the expression of an oncogenic transcription factor called E2A-PBX1 in cases of acute leukemia. In the work described here, we elucidate the interaction between E2A-PBX1 and transcriptional co-activators. We confirm that the E2A portion can interact with CBP and PCAF and map required elements on E2A and CBP. On CBP, the interaction involves the KIX domain, a well-characterized domain that mediates interactions with several other oncogenic transcription factors. On E2A, the interaction with CBP requires conserved a-helical domains that reside within activation domains 1 and 2 (AD1 and AD2, respectively). Using purified, recombinant proteins, we show that the E2A-CBP interaction is direct. Notwithstanding the previously demonstrated ability of AD1 and AD2 to function independently, some of our findings suggest functional cooperativity between these two domains. Finally, we show that the CBP/p300-interactive helical domains of E2A are important in the induction of proliferation in cultured primary bone marrow cells retrovirally transduced with E2A-PBX1. Our findings suggest that some aspects of E2A-PBX1 oncogenesis involve a direct interaction with the KIX domain of CBP/p300.


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