Characterization of the hematopoietic transcription factor NF-E2 in primary murine megakaryocytes

J Biol Chem. 1998 Mar 27;273(13):7572-8. doi: 10.1074/jbc.273.13.7572.


Biochemical analysis of megakaryocytes, the precursors of blood platelets, is limited by their rarity in vivo, and studies on lineage-specific gene expression have been conducted exclusively in cell lines with limited megakaryocytic potential. Mice lacking the transcription factor NF-E2 display arrested megakaryocyte differentiation and profound thrombocytopenia. To study the heterodimeric NF-E2 protein in primary cells, we cultured mouse fetal livers with the c-Mpl ligand, obtained highly enriched megakaryocyte populations, and readily detected NF-E2 activity in nuclear extracts. As in erythroid cells, p45 NF-E2 is the only large subunit in primary megakaryocytes that dimerizes with distinct small Maf proteins to constitute a heterogeneous NF-E2 complex. Whereas p18/MafK is the predominant small Maf protein in erythroid cells, the related polypeptides MafG and/or MafF predominate in megakaryocytes. Although this represents the first example of differential small Maf protein expression among closely related blood lineages, the DNA-binding specificity of NF-E2 is similar in both cell types. Although the megakaryocyte protein preferentially binds an asymmetric AP-1-related motif, it also recognizes cAMP-responsive element-related sequences, albeit with lower affinity, and nucleotides outside the core sequence influence the DNA-protein interaction. These results demonstrate the feasibility of biochemical studies on primary murine megakaryocytes and provide a basis to dissect the critical functions of NF-E2 in megakaryocyte differentiation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Cell Differentiation
  • Culture Techniques
  • DNA / chemistry
  • DNA / metabolism
  • DNA-Binding Proteins / metabolism*
  • Erythroid-Specific DNA-Binding Factors
  • Liver / embryology
  • MafK Transcription Factor
  • Megakaryocytes / cytology
  • Megakaryocytes / metabolism*
  • Mice
  • NF-E2 Transcription Factor
  • NF-E2 Transcription Factor, p45 Subunit
  • Nuclear Proteins / metabolism*
  • Sequence Analysis, DNA
  • Thrombopoietin / metabolism
  • Transcription Factors / metabolism*
  • Tumor Cells, Cultured
  • Zinc Fingers*


  • DNA-Binding Proteins
  • Erythroid-Specific DNA-Binding Factors
  • MafK Transcription Factor
  • Mafk protein, mouse
  • NF-E2 Transcription Factor
  • NF-E2 Transcription Factor, p45 Subunit
  • Nfe2 protein, mouse
  • Nuclear Proteins
  • Transcription Factors
  • DNA
  • Thrombopoietin