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.

Distinct Cytoplasmic Regions of the Human Granulocyte Colony-Stimulating Factor Receptor Involved in Induction of Proliferation and Maturation

“…It is likely that this region is involved in activation of common or highly related signal transduction pathways. This notion is supported by several receptor transfection studies (7,15,29,35,48) and by the fact that some of the ligands for these receptors induce similar patterns of tyrosine phosphorylation (9,27,36).…”

Protein kinase C-dependent phosphorylation is involved in resistance to tumour necrosis factor-α-induced cytotoxicity in a human monocytoid cell line

“…Although TNFa receptors do not themselves appear to have intrinsic kinase activity (Schall et al, 1990;Smith et al, 1990), several reports (Schutze et al, 1989;Kaur et al, 1989;Robaye et al, 1989;Evans et al, 1990) have described the TNFa-induced phosphorylation of distinct cytosolic proteins in different human of molecular masses 130, 90, 80, 65 and 42 kDa was found. Subsequently, we found a similar pattern of increased phosphorylation following stimulation of AlO cells with mezerein, a phorbol ester derivative which activates PKC, a serine/threonine kinase.…”

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).…”