Binding of G-CSF, GM-CSF, tumor necrosis factor-alpha, and gamma- interferon to cell surface receptors on human myeloid leukemia cells triggers rapid tyrosine and serine phosphorylation of a 75-Kd protein

Abstract

Granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), gamma-interferon (gamma-IFN), or tumor necrosis factor-alpha (TNF-alpha) triggered the rapid, stable phosphorylation of a 75-Kd protein (p75) when incubated with permeabilized HL60 human myeloid leukemia cells in the presence of [gamma-32P] ATP. Among several chemical inducers of HL60 cell differentiation, dimethyl sulfoxide also triggered p75 labeling, but retinoic acid or 12-O-tetradecanoylphorbol-13-acetate did not elicit this response. Pretreatment of cells with G-CSF or GM-CSF for more than 30 seconds before permeabilization rendered the p75 labeling undetectable, suggesting that ligand-stimulated labeling was rapidly completed within this time in intact cells. Phosphorylation of p75 occurred on serine and tyrosine residues. This conclusion was confirmed by direct phosphoamino acid analysis. Immunoblot analysis of lysates of intact HL60 cells that had been incubated with G-CSF, GM-CSF, IFN, or TNF confirmed that tyrosine phosphorylation of a p75 also occurred in response to these cytokines in intact cells. Pretreatment of intact HL60 cells with one biologic agent or dimethyl sulfoxide abolished p75 labeling in response to incubation of permeabilized cells with a second agent, strongly suggesting that the same protein was phosphorylated in response to these treatments. p75 labeling was strictly dependent on expression of the appropriate ligand receptor. Data suggest that activation of a tyrosine kinase system is an early response to the binding of G-CSF, GM-CSF, TNF, or IFN to their respective cell surface receptors, or to the addition of dimethyl sulfoxide, and that the resulting phosphorylation event(s) may play a role in securing common elements in the biologic responses to these agents.

Tumor necrosis factor induces rapid production of 1'2'diacylglycerol by a phosphatidylcholine-specific phospholipase C.

“…AA can be metabolized to leukotrienes and prostaglandins, potent mediators of inflammatory processes. In addition, protein kinases (PK) appear to play a crucial role in TNF signal transduction pathways (27)(28)(29)(30)(31), and a number of cellular substrates for TNF-responsive PK have been detected (32)(33)(34)(35)(36)(37)(38)(39). We have recently shown that TNF stimulates translocation and activation of PKC (30).…”

Stimulation of the metastatic properties of lewis‐lung‐carcinoma cells by autologous granulocyte‐macrophage colony‐stimulating factor

“…GM-CSF stimulates kinase activity in normal and in tumor cells (Gomez-Cambronero et aL, 1989;Evans et al, 1990). In addition, migration of LLC-LN7 is stimulated upon PKA activation, but is inhibited upon PKC activation (Young et al,1 9 9 1~) .…”

GM‐CSF triggers a rapid, glucose dependent extracellular acidification by TF‐1 cells: Evidence for sodium/proton antiporter and PKC mediated activation of acid production

“…In fact, Kitamura et al (1991b) demonstrated that hIL-3 and hGM-CSF receptors share a common p-subunit by cloning c-DNA for the hIL-3 receptor a-subunit, which reconstituted the high-affinity IL-3 receptor when co-transfected with the hGM-CSF receptor P-subunit. Since there is evidence for early GM-CSF and IL-3 induced tyrosine phosphorylation in hemopoetic cells (Isfort and Ihle, 1990;Evans et al, 1990) and no consensus tyrosine kinase sequences were found in the cloned receptor subunits (Hayashida et al,199Oj,it has been suggested that additional signal transducing elements are associated with the IL-3IGM-CSF receptor complex.…”