PU.1 activation relieves GATA-1-mediated repression of Cebpa and Cbfb during leukemia differentiation

Pavel Burda, Nikola Curik, Juraj Kokavec, Petra Basova, Dana Mikulenkova, Arthur I. Skoultchi, Jiri Zavadil, Tomas Stopka

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Hematopoietic transcription factors GATA-1 and PU.1 bind each other on DNA to block transcriptional programs of undesired lineage during hematopoietic commitment. Murine erythroleukemia (MEL) cells that coexpress GATA-1 and PU.1 are blocked at the blast stage but respond to molecular removal (downregulation) of PU.1 or addition (upregulation) of GATA-1 by inducing terminal erythroid differentiation. To test whether GATA-1 blocks PU.1 in MEL cells,we have conditionally activated a transgenic PU.1 protein fused with the estrogen receptor ligand-binding domain (PUER),resulting in activation of a myeloid transcriptional program. Gene expression arrays identified components of the PU.1-dependent transcriptome negatively regulated by GATA-1 in MEL cells,including CCAAT/enhancer binding protein á (Cebpa) and core-binding factor, â subunit (Cbfb),which encode two key hematopoietic transcription factors. Inhibition of GATA-1 by small interfering RNA resulted in derepression of PU.1 target genes. Chromatin immunoprecipitation and reporter assays identified PU.1 motif sequences near Cebpa and Cbfb that are co-occupied by PU.1 and GATA-1 in the leukemic blasts. Significant derepression of Cebpa and Cbfb is achieved in MEL cells by either activation of PU.1 or knockdown of GATA-1. Furthermore,transcriptional regulation of these loci by manipulating the levels of PU.1 and GATA-1 involves quantitative increases in a transcriptionally active chromatin mark: acetylation of histone H3K9. Collectively,we show that either activation of PU.1 or inhibition of GATA-1 efficiently reverses the transcriptional block imposed by GATA-1 and leads to the activation of a myeloid transcriptional program directed by PU.1.

Original languageEnglish (US)
Pages (from-to)1693-1703
Number of pages11
JournalMolecular Cancer Research
Volume7
Issue number10
DOIs
StatePublished - Oct 2009

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Core Binding Factors
Leukemia, Erythroblastic, Acute
Leukemia
Histone Code
CCAAT-Enhancer-Binding Proteins
Chromatin Immunoprecipitation
Acetylation
Transcriptome
Estrogen Receptors
Small Interfering RNA
Chromatin
Transcription Factors
Up-Regulation
Down-Regulation
Ligands
Gene Expression
DNA
Genes
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Oncology

Cite this

PU.1 activation relieves GATA-1-mediated repression of Cebpa and Cbfb during leukemia differentiation. / Burda, Pavel; Curik, Nikola; Kokavec, Juraj; Basova, Petra; Mikulenkova, Dana; Skoultchi, Arthur I.; Zavadil, Jiri; Stopka, Tomas.

In: Molecular Cancer Research, Vol. 7, No. 10, 10.2009, p. 1693-1703.

Research output: Contribution to journalArticle

Burda, P, Curik, N, Kokavec, J, Basova, P, Mikulenkova, D, Skoultchi, AI, Zavadil, J & Stopka, T 2009, 'PU.1 activation relieves GATA-1-mediated repression of Cebpa and Cbfb during leukemia differentiation', Molecular Cancer Research, vol. 7, no. 10, pp. 1693-1703. https://doi.org/10.1158/1541-7786.MCR-09-0031
Burda, Pavel ; Curik, Nikola ; Kokavec, Juraj ; Basova, Petra ; Mikulenkova, Dana ; Skoultchi, Arthur I. ; Zavadil, Jiri ; Stopka, Tomas. / PU.1 activation relieves GATA-1-mediated repression of Cebpa and Cbfb during leukemia differentiation. In: Molecular Cancer Research. 2009 ; Vol. 7, No. 10. pp. 1693-1703.
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AU - Mikulenkova, Dana

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AU - Stopka, Tomas

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AB - Hematopoietic transcription factors GATA-1 and PU.1 bind each other on DNA to block transcriptional programs of undesired lineage during hematopoietic commitment. Murine erythroleukemia (MEL) cells that coexpress GATA-1 and PU.1 are blocked at the blast stage but respond to molecular removal (downregulation) of PU.1 or addition (upregulation) of GATA-1 by inducing terminal erythroid differentiation. To test whether GATA-1 blocks PU.1 in MEL cells,we have conditionally activated a transgenic PU.1 protein fused with the estrogen receptor ligand-binding domain (PUER),resulting in activation of a myeloid transcriptional program. Gene expression arrays identified components of the PU.1-dependent transcriptome negatively regulated by GATA-1 in MEL cells,including CCAAT/enhancer binding protein á (Cebpa) and core-binding factor, â subunit (Cbfb),which encode two key hematopoietic transcription factors. Inhibition of GATA-1 by small interfering RNA resulted in derepression of PU.1 target genes. Chromatin immunoprecipitation and reporter assays identified PU.1 motif sequences near Cebpa and Cbfb that are co-occupied by PU.1 and GATA-1 in the leukemic blasts. Significant derepression of Cebpa and Cbfb is achieved in MEL cells by either activation of PU.1 or knockdown of GATA-1. Furthermore,transcriptional regulation of these loci by manipulating the levels of PU.1 and GATA-1 involves quantitative increases in a transcriptionally active chromatin mark: acetylation of histone H3K9. Collectively,we show that either activation of PU.1 or inhibition of GATA-1 efficiently reverses the transcriptional block imposed by GATA-1 and leads to the activation of a myeloid transcriptional program directed by PU.1.

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