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Interleukin-3 signals through multiple isoforms of Stat5

M Azam1, H Erdjument-Bromage, B L Kreider

  • 1Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.

The EMBO Journal
|April 3, 1995
PubMed
Summary

Interleukin-3 (IL-3) activates distinct signal transducing factors (STFs) in myeloid cells. These factors, identified as Stat5 isoforms, regulate myeloid growth and maturation by binding to related receptors.

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Area of Science:

  • Cellular signaling pathways
  • Hematopoiesis and myeloid cell development
  • Cytokine receptor interactions

Background:

  • Interleukin-3 (IL-3) family cytokines are crucial for myeloid cell growth and maturation.
  • These cytokines exert effects by binding to specific receptors, initiating intracellular signaling cascades.
  • Recent findings suggest the involvement of cytoplasmic signal transducing factors (STFs) in mediating IL-3's effects, particularly the induction of immediate early genes.

Purpose of the Study:

  • To identify and characterize the cytoplasmic signal transducing factors (STFs) activated by IL-3 in myeloid cells.
  • To elucidate the molecular composition and relationships between STFs activated in immature versus mature myeloid cells.
  • To determine if these STFs are related to known signaling proteins and to investigate their role in IL-3-mediated signaling.

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Main Methods:

  • Purification and biochemical analysis of tyrosine-phosphorylated DNA binding proteins (77 and 80 kDa) from IL-3-stimulated immature myeloid cells.
  • Peptide sequencing and nucleotide sequence analysis to determine protein identity and gene origin.
  • Generation of antiserum against purified proteins to probe for related proteins in mature myeloid cells.
  • Stimulation of mature myeloid cells with IL-3 and granulocyte-macrophage colony-stimulating factor (GM-CSF) and analysis of activated STFs.

Main Results:

  • IL-3 activates STF-IL-3a (77 and 80 kDa proteins) in immature myeloid cells and STF-IL-3b (94 and 96 kDa proteins) in mature myeloid cells.
  • The 77 and 80 kDa proteins are identified as murine homologs of ovine mammary gland factor (MGF)/Stat5, encoded by distinct genes.
  • Phosphorylation of p77 and p80 occurs at tyrosine 699, analogous to interferon-stimulated signaling.
  • Antiserum against p77/p80 recognizes the components of STF-IL-3b, suggesting they are also Stat5 isoforms.

Conclusions:

  • The IL-3 cytokine family activates multiple isoforms of the signal transducing protein Stat5.
  • These Stat5 isoforms play a critical role in mediating the effects of IL-3 on myeloid cell growth, maturation, and gene expression.
  • The findings reveal a conserved mechanism of cytokine signaling involving Stat5 activation across different myeloid cell types and stimuli.