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Mol. Cell. Biol., Jan 1998, 322-333, Vol 18, No. 1
Copyright © 1998, American Society for Microbiology
The t(8;21) fusion product, AML-1-ETO, associates with C/EBP-alpha, inhibits C/EBP-alpha-dependent transcription, and blocks granulocytic differentiation
JJ Westendorf, CM Yamamoto, N Lenny, JR Downing, ME Selsted and SW Hiebert
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
AML-1B is a hematopoietic transcription factor that is functionally
inactivated by multiple chromosomal translocations in human acute
myeloblastic and B-cell lymphocytic leukemias. The t(8;21)(q22;q22)
translocation replaces the C terminus, including the transactivation domain
of AML-1B, with ETO, a nuclear protein of unknown function. We previously
showed that AML-1-ETO is a dominant inhibitor of AML-1B- dependent
transcriptional activation. Here we demonstrate that AML-1- ETO also
inhibits C/EBP-alpha-dependent activation of the myeloid cell- specific,
rat defensin NP-3 promoter. AML-1B bound the core enhancer motifs present
in the NP-3 promoter and activated transcription approximately sixfold.
Similarly, C/EBP-alpha bound NP-3 promoter sequences and activated
transcription approximately sixfold. Coexpression of C/EBP-alpha with
AML-1B or its family members, AML-2 and murine AML-3, synergistically
activated the NP-3 promoter up to 60- fold. The t(8;21) product, AML-1-ETO,
repressed AML-1B-dependent activation of NP-3 and completely inhibited
C/EBP-alpha-dependent activity as well as the synergistic activation. In
contrast, the inv(16) product, which indirectly targets AML family members
by fusing their heterodimeric DNA binding partner, CBF-beta, to the myosin
heavy chain, inhibited AML-1B but not C/EBP-alpha activation or the
synergistic activation. AML-1-ETO and C/EBP-alpha were coimmunoprecipitated
and thus physically interact in vivo. Deletion mutants demonstrated that
the C terminus of ETO was required for AML-1- ETO-mediated repression of
the synergistic activation but not for association with C/EBP-alpha.
Finally, overexpression of AML-1-ETO in myeloid progenitor cells prevented
granulocyte colony-stimulating factor-induced differentiation. Thus,
AML-1-ETO may contribute to leukemogenesis by specifically inhibiting
C/EBP-alpha- and AML-1B- dependent activation of myeloid promoters and
blocking differentiation.
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