Transcriptional regulation of myeloid differentiation primary response (MyD) genes during myeloid differentiation is mediated by nuclear factor Y

Blood. 2002 Jul 1;100(1):80-8. doi: 10.1182/blood.v100.1.80.

Abstract

To understand the molecular mechanism by which interleukin-6 (IL-6) regulates myeloid differentiation primary response (MyD) genes at the onset of M1 myeloid differentiation, we used JunB as a representative MyD gene to isolate and characterize IL-6 responsive elements. An IL-6 responsive element was localized between -65 and -52 of the JunB promoter (-65/-52 IL-6RE). By using antibody and oligonucleotide competition assays in electrophoretic mobility shift assay experiments, we have shown that the heterotrimeric transcription nuclear factor Y (NF-Y) complex binds to this element. A dominant-negative form of NF-YA, ectopically expressed in M1 cells, blocked NF-Y binding to the -65/-52 IL-6RE and reduced induction of JunB by IL-6. Furthermore, inhibition of NF-Y binding also reduced MyD gene induction by IL-6 and dampened the IL-6-induced M1 differentiation program. These findings are consistent with the observation that most MyD genes contain intact NF-Y binding motifs in their promoter regions. In contrast to M1 cells, during myeloid differentiation of bone marrow (BM), there was induction of NF-Y binding to the -65/-52 IL-6RE. This induced binding can be attributed to the observed induction of NF-YA protein expression and may reflect the molecular mechanism that couples proliferation to terminal differentiation of normal myeloblasts. Similar to M1 cells, blocking NF-Y binding in BM resulted in a reduction in mature macrophages. It can be concluded that NF-Y plays a role in the transcriptional regulation of MyD genes and is required for optimum myeloid differentiation.

Publication types

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

MeSH terms

  • Animals
  • Bone Marrow Cells
  • CCAAT-Binding Factor / metabolism
  • CCAAT-Binding Factor / pharmacology
  • CCAAT-Binding Factor / physiology*
  • Cell Differentiation / drug effects
  • Cell Differentiation / genetics
  • Drug Interactions
  • Female
  • Gene Expression Regulation / drug effects*
  • Genes, jun / drug effects
  • Interleukin-6 / genetics
  • Interleukin-6 / pharmacology
  • Mice
  • Mice, Inbred BALB C
  • Myeloid Cells / cytology*
  • Myeloid Cells / metabolism
  • Protein Binding
  • Response Elements
  • Transcription Factors / drug effects
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Transcription Factors / pharmacology
  • Transcription Factors / physiology*
  • Transcriptional Activation
  • Tumor Cells, Cultured

Substances

  • CCAAT-Binding Factor
  • Interleukin-6
  • Transcription Factors
  • nuclear factor Y