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Granulocyte-macrophage colony-stimulating factor-activated signaling
pathways in human neutrophils. I. Tyrosine phosphorylation-dependent
stimulation of phosphatidylinositol 3-kinase and inhibition by phorbol
esters
A al-Shami, SG Bourgoin and PH Naccache
Centre de Recherche en Rhumatologie et Immunologie, Faculty of Medicine,
Laval University, Ste-Foy, Quebec, Canada.
Phosphatidylinositol 3-kinase (PI3-kinase) is a cytosolic enzyme that plays
key roles in mediating signaling through many receptors. The heterodimeric
form of PI3-kinase is made up of a regulatory subunit, p85, and a catalytic
subunit, p110. Although granulocyte-macrophage colony-stimulating factor
(GM-CSF) has been shown to activate PI3- kinase, the mechanisms by which
this activation is mediated and regulated are incompletely understood. Here
we show that treatment of human neutrophils with GM-CSF induced both time-
and concentration- dependent increases in the level of tyrosine
phosphorylation of p85. The ability of GM-CSF to activate PI3-kinase was
abolished by pretreating the cells with erbstatin, a tyrosine kinase
inhibitor. The simultaneous treatment of the cells with GM-CSF and phorbol
esters such as phorbol 12-myristate 13-acetate (PMA) and phorbol
12,13-dibutyrate (PDBu) significantly inhibited both the tyrosine
phosphorylation of p85 and the activation of PI3-kinase. The inhibitory
effects of phorbol esters were not induced by their inactive analogues and
they were selective to the stimulation of tyrosine phosphorylation of p85
since phorbol esters did not alter the enhancement of the pattern of
tyrosine phosphorylation of other cellular proteins, including that of Jak2
induced by GM-CSF. However, PMA significantly inhibited the in situ
tyrosine phosphorylation and the activation of lyn observed in response to
GM-CSF. The results suggest that the activation of PI3-kinase by GM- CSF is
mediated by the tyrosine phosphorylation of p85 and that this activation is
downregulated by PKC possibly via the inhibition of lyn.
Volume 89,
Issue 3,
pp. 1035-1044,
02/01/1997
Copyright © 1997 by The American Society of Hematology
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