GATA factors are essential for transcription of the survival gene E4bp4 and the viability response of interleukin-3 in Ba/F3 hematopoietic cells

J Biol Chem. 2002 Jul 26;277(30):27144-53. doi: 10.1074/jbc.M200924200. Epub 2002 May 21.

Abstract

E4bp4, a member of the basic region/leucine zipper transcription factor superfamily, is up-regulated by the interleukin-3 (IL-3) signaling pathway and plays an important role in the anti-apoptotic response of IL-3. In this study, we demonstrated that E4bp4 is regulated by IL-3 mainly at the transcriptional level. Promoter analysis revealed that a GATA motif downstream of a major transcription initiation site is essential for E4bp4 expression in the IL-3-dependent Ba/F3 cell line. Gel shift assays demonstrated that both GATA-1 and GATA-2 proteins bind to the E4bp4 GATA site in vitro, and the chromatin immunoprecipitation assay further confirmed the in vivo binding of GATA-1 to the E4bp4 promoter. Overexpression of GATA-1 alone transactivates the E4bp4 reporter, whereas transactivation of the E4bp4 reporter by GATA-2 is dependent on the stimulation of IL-3. Last, we demonstrated that alteration of GATA-1 binding to the GATA site by stably overexpressing GATA-1 or a GATA-1 mutant containing only the DNA-binding domain not only modulates the expression of the E4bp4 gene but also influences apoptosis induced by IL-3 removal. Taken together, our results suggest that the GATA factors play an important role in transducing the survival signal of IL-3, and one of their cellular targets is E4bp4.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Base Sequence
  • Basic-Leucine Zipper Transcription Factors
  • Blotting, Northern
  • Blotting, Western
  • Cell Survival
  • Chromatin / metabolism
  • Chromosome Mapping
  • Cloning, Molecular
  • DNA / metabolism
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • DNA-Binding Proteins / physiology*
  • Electroporation
  • Erythroid-Specific DNA-Binding Factors
  • G-Box Binding Factors
  • GATA1 Transcription Factor
  • GATA2 Transcription Factor
  • Genes, Reporter
  • Hematopoietic Stem Cells / metabolism
  • Interleukin-3 / metabolism*
  • Luciferases / metabolism
  • Mice
  • Models, Genetic
  • Molecular Sequence Data
  • Plasmids / metabolism
  • Precipitin Tests
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA, Messenger / metabolism
  • Signal Transduction
  • Single-Strand Specific DNA and RNA Endonucleases / metabolism
  • Time Factors
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcription Factors / physiology*
  • Transcription, Genetic*
  • Transcriptional Activation

Substances

  • Basic-Leucine Zipper Transcription Factors
  • Chromatin
  • DNA-Binding Proteins
  • Erythroid-Specific DNA-Binding Factors
  • G-Box Binding Factors
  • GATA1 Transcription Factor
  • GATA2 Transcription Factor
  • Gata1 protein, mouse
  • Gata2 protein, mouse
  • Interleukin-3
  • Nfil3 protein, mouse
  • RNA, Messenger
  • Transcription Factors
  • DNA
  • Luciferases
  • Single-Strand Specific DNA and RNA Endonucleases