Granulocyte colony-stimulating factor activates protein kinase A in granulocytic but not monocytic precursors or neutrophils.

Granulocyte colony-stimulating factor (G-CSF) regulates survival, proliferation, differentiation, and activation of myeloid cells. G-CSF-R signaling mechanisms other than tyrosine kinase activation have not been documented. We explored the potential involvement of cAMP-dependent protein kinase A (PKA) in G-CSF-R signal transduction. In this report, we provide the first direct evidence of PKA modulation by G-CSF-R. G-CSF treatment of granulocytic precursor cell lines (HL-60, NFS-60, KG-1) resulted in PKA activation, measured by phosphorylation of Kemptide, a peptide substrate. In contrast, the myelomonocytic cell lines (WEHI-3B,U-937) and peripheral blood neutrophils (PMNC) showed a rapid decrease in PKA activity in response to G-CSF. H-89, a specific inhibitor of PKA, blocked G-CSF-induced PKA activation in HL-60 cells but did not affect ligand-induced downmodulation of G-CSF-R. Indomethacin, an inhibitor of the cyclooxygenase pathway and prostaglandin synthesis, did not inhibit PKA induction in G-CSF-treated HL-60 cells. Our results demonstrate the involvement of PKA in G-CSF-R signal transduction and suggest a lineage-restricted, developmental stage-specific regulation of this pathway in myeloid cells.