Abstract
The heterodimeric transcription factor RUNX1/PEBP2-beta (also known as AML1/CBF-beta) is essential for definitive hematopoiesis. Here, we show that interaction with PEBP2-beta leads to the phosphorylation of RUNX1, which in turn induces p300 phosphorylation. This is mediated by homeodomain interacting kinase 2 (HIPK2), targeting Ser(249), Ser(273), and Thr(276) in RUNX1, in a manner that is also dependent on the RUNX1 PY motif. Importantly, we observed the in vitro disruption of this phosphorylation cascade by multiple leukemogenic genetic defects targeting RUNX1/CBFB. In particular, the oncogenic protein PEBP2-beta-SMMHC prevents RUNX1/p300 phosphorylation by sequestering HIPK2 to mislocalized RUNX1/beta-SMMHC complexes. Therefore, phosphorylation of RUNX1 appears a critical step in its association with and phosphorylation of p300, and its disruption may be a common theme in RUNX1-associated leukemogenesis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Base Sequence
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CCAAT-Binding Factor / metabolism*
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COS Cells
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Carrier Proteins / metabolism*
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Cell Line, Tumor
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Chlorocebus aethiops
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Core Binding Factor Alpha 2 Subunit / chemistry
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Core Binding Factor Alpha 2 Subunit / genetics
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Core Binding Factor Alpha 2 Subunit / metabolism*
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Core Binding Factor beta Subunit / chemistry
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Core Binding Factor beta Subunit / metabolism*
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DNA Primers / genetics
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Humans
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K562 Cells
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Leukemia / etiology*
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Leukemia / genetics
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Leukemia / metabolism
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Mice
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Multiprotein Complexes
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Mutagenesis, Site-Directed
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Phosphorylation
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Protein Serine-Threonine Kinases / metabolism*
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Protein Structure, Tertiary
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Deletion
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p300-CBP Transcription Factors / chemistry
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p300-CBP Transcription Factors / metabolism*
Substances
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CBFB protein, human
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CCAAT-Binding Factor
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Carrier Proteins
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Core Binding Factor Alpha 2 Subunit
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Core Binding Factor beta Subunit
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DNA Primers
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Multiprotein Complexes
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RUNX1 protein, human
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Recombinant Proteins
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p300-CBP Transcription Factors
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HIPK2 protein, human
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Protein Serine-Threonine Kinases