Phorbol ester treatment of K562 cells regulates the transcriptional activity of AML1c through phosphorylation

Youhong Zhang, Joseph R. Biggs, Andrew Kraft

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We find that phorbol ester (PE) treatment of K562 cells greatly stimulates promoters (T cell receptor β, myeloperoxidase, macrophage colony-stimulating factor receptor, and granulocyte macrophage colony-stimulating factor receptor) containing AML1 transcription factor binding sites. This stimulation of AML1c transcriptional activity is mediated by direct phosphorylation of the AML1c molecule on multiple phosphorylation sites. Eleven AML1c (S/T)P sites in the transcriptional activating domain are phosphorylated at a basal level in untreated K562 cells; treatment of the K562 cells with PE results in increased phosphorylation at five of these sites (serines 276, 293, 303, 462, and threonine 300). Mutation of these five sites to alanine inhibits PE-induced transcriptional activity; mutation of the sites to an acidic amino acid, aspartic acid, stimulates constitutive activity. Single mutations in four amino acids or double mutations (serines 273 and 293 or threonine 300 and serine 303) have little effect on AML1c transcriptional activity. Inhibitor assays suggest that the ERK family of protein kinases is activated by PEs to phosphorylate the (S/T)P sites within the AML1c molecule and markedly enhance the transcriptional activity of AML1c.

Original languageEnglish (US)
Pages (from-to)53116-53125
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number51
DOIs
StatePublished - Dec 17 2004
Externally publishedYes

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Phosphorylation
K562 Cells
Phorbol Esters
Threonine
Serine
Mutation
Granulocyte-Macrophage Colony-Stimulating Factor Receptors
Macrophage Colony-Stimulating Factor Receptors
Acidic Amino Acids
Molecules
Macrophages
T-Cell Antigen Receptor
Aspartic Acid
Alanine
Protein Kinases
Peroxidase
Assays
Transcription Factors
Amino Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phorbol ester treatment of K562 cells regulates the transcriptional activity of AML1c through phosphorylation. / Zhang, Youhong; Biggs, Joseph R.; Kraft, Andrew.

In: Journal of Biological Chemistry, Vol. 279, No. 51, 17.12.2004, p. 53116-53125.

Research output: Contribution to journalArticle

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