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Phosphorylation of RUNX1 by cyclin-dependent kinase reduces direct interaction with HDAC1 and HDAC3

Hong Guo1, Alan D Friedman

  • 1Division of Pediatric Oncology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USA.

The Journal of Biological Chemistry
|November 10, 2010
PubMed

Insights

RUNX1 phosphorylation by cyclin-dependent kinases (cdks) reduces its interaction with HDAC1/3, enhancing hematopoietic stem cell proliferation. This suggests a mechanism for RUNX1 in leukemic transformation and potential therapeutic targeting.

Area of Science:

  • Hematology
  • Molecular Biology
  • Cancer Research

Background:

  • RUNX1 is crucial for hematopoietic stem cell development and lineage maturation.
  • RUNX1 mutations are implicated in leukemic transformation.
  • Cyclin-dependent kinases (cdks) phosphorylate RUNX1, enhancing its trans-activation activity.

Purpose of the Study:

  • To investigate the interaction between RUNX1 and histone deacetylases (HDACs).
  • To determine the role of RUNX1 phosphorylation sites (Ser-48, Ser-303, Ser-424) in HDAC binding.
  • To elucidate the functional consequences of altered RUNX1-HDAC interaction on hematopoietic progenitor cell proliferation.

Main Methods:

  • Co-immunoprecipitation assays to assess RUNX1-HDAC1/3 interactions in cells and in vitro.
  • Site-directed mutagenesis of RUNX1 phosphorylation sites (Ser to Asp or Ala).
  • In vitro phosphorylation of RUNX1 using cdk1/cyclinB.
  • Assessment of hematopoietic progenitor cell proliferation following transduction with modified RUNX1.

Main Results:

  • Endogenous RUNX1 interacts with HDAC1 and HDAC3.
  • Mutation of RUNX1 serines to aspartic acid reduced HDAC interaction, while mutation to alanine increased it.
  • RUNX1 phosphorylation by cdks, mimicking dephosphorylation at these sites, reduced HDAC binding.
  • RUNX1 variants that reduced HDAC interaction stimulated marrow progenitor cell proliferation more potently.

Conclusions:

  • RUNX1 phosphorylation by cdks decreases its interaction with HDAC1/3.
  • Reduced RUNX1-HDAC interaction enhances RUNX1 trans-activation and stimulates hematopoietic progenitor cell proliferation.
  • This mechanism provides insight into RUNX1's role in hematopoiesis and leukemogenesis.

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