Acetylation of C/EBPε is a prerequisite for terminal neutrophil differentiation

Blood. 2015 Mar 12;125(11):1782-92. doi: 10.1182/blood-2013-12-543850. Epub 2015 Jan 7.

Abstract

C/EBPε, a member of the CCAAT/enhancer binding protein (C/EBP) family of transcription factors, is exclusively expressed in myeloid cells and regulates transition from the promyelocytic stage to the myelocytic stage of neutrophil development, being indispensable for secondary and tertiary granule formation. Knowledge concerning the functional role of C/EBPε posttranslational modifications is limited to studies concerning phosphorylation and sumoylation. In the current study, using ectopic expression and ex vivo differentiation of CD34(+) hematopoietic progenitor cells, we demonstrate that C/EBPε is acetylated, which was confirmed by mass spectrometry analysis, identifying 4 acetylated lysines in 3 distinct functional domains. Regulation of C/EBPε acetylation levels by the p300 acetyltransferase and the sirtuin 1 deacetylase controls transcriptional activity, which can at least in part be explained by modulation of DNA binding. During neutrophil development, acetylation of lysines 121 and 198 were found to be crucial for terminal neutrophil differentiation and the expression of neutrophil-specific granule proteins, including lactoferrin and collagenase. Taken together, our data illustrate a critical role for acetylation in the functional regulation of C/EBPε activity during terminal neutrophil development.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Animals
  • CCAAT-Enhancer-Binding Proteins / chemistry
  • CCAAT-Enhancer-Binding Proteins / genetics
  • CCAAT-Enhancer-Binding Proteins / metabolism*
  • COS Cells
  • Cell Differentiation
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Collagenases / metabolism
  • HL-60 Cells
  • Humans
  • Lactoferrin / metabolism
  • Lysine / chemistry
  • Myelopoiesis
  • Neutrophils / cytology*
  • Neutrophils / metabolism*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sirtuin 1 / metabolism
  • Transcription, Genetic
  • p300-CBP Transcription Factors / metabolism

Substances

  • CCAAT-Enhancer-Binding Proteins
  • LTF protein, human
  • Recombinant Fusion Proteins
  • CEBPE protein, human
  • p300-CBP Transcription Factors
  • p300-CBP-associated factor
  • Lactoferrin
  • Collagenases
  • SIRT1 protein, human
  • Sirtuin 1
  • Lysine