Gene activation requires cooperative assembly of multiprotein
transcription factor-coregulator complexes. Disruption to cooperative assemblage could underlie repression of tumor suppressor genes in
leukemia cells. Mechanisms of cooperation and its disruption were therefore examined for PU.1 and RUNX1,
transcription factors that cooperate to activate hematopoietic differentiation genes. PU.1 is highly expressed in
leukemia cells, whereas RUNX1 is frequently inactivated by mutation or translocation. Thus, coregulator interactions of Pu.1 were examined by immunoprecipitation coupled with tandem mass spectrometry/Western blot in wild-type and Runx1-deficient hematopoietic cells. In wild-type cells, the NuAT and Baf families of coactivators coimmunoprecipitated with Pu.1. Runx1 deficiency produced a striking switch to Pu.1 interaction with the Dnmt1, Sin3A, Nurd, CoRest, and B-Wich
corepressor families.
Corepressors of the Polycomb family, which are frequently inactivated by mutation or deletion in
myeloid leukemia, did not interact with Pu.1. The most significant gene ontology association of Runx1-Pu.1 co-bound genes was with macrophages, therefore, functional consequences of altered
corepressor/coactivator exchange were examined at Mcsfr, a key macrophage differentiation gene. In
chromatin immunoprecipitation analyses, high level Pu.1 binding to the Mcsfr promoter was not decreased by Runx1 deficiency. However, the Pu.1-driven shift from
histone repression to activation marks at this locus, and terminal macrophage differentiation, were substantially diminished. DNMT1 inhibition, but not Polycomb inhibition, in RUNX1-translocated
leukemia cells induced terminal differentiation. Thus, RUNX1 and PU.1 cooperate to exchange
corepressors for coactivators, and the specific
corepressors recruited to PU.1 as a consequence of RUNX1 deficiency could be rational targets for
leukemia differentiation
therapy.