Involvement of tyrosine kinase and protein kinase C in platelet-activating-factor-induced c-fos gene expression in A-431 cells

Y. B. Tripathi, R. W. Lim, S. Fernandez-Gallardo, J. C. Kandala, Ramareddy Guntaka, S. D. Shukla

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Abstract

In A-431 cells, platelet-activating factor (PAF) induces the expression of c-fos and TIS-1 genes in both the absence and the presence of cycloheximide in a structurally specific and receptor-coupled manner. We have now investigated the molecular mechanisms of this response, particularly in relation to the role of protein kinases. Pretreatment of cells with genistein or methyl-2,5-dihydroxycinnamate (tyrosine kinase inhibitors) or staurosporine (a protein kinase C inhibitor) for 20min abolished the c-fos expression induced by PAF. Interestingly when genistein was added 90 s after addition of PAF, no inhibition was observed. Similarly. staurosporine did not inhibit c-fos expression when added 8 min after PAF addition to the cells. These inhibitions were dose-dependent (IC50 for staurosporine was 180 nM, and for genistein 50 μM). Simultaneous addition of PAF and phorbol 12-myristate 13-acetate (PMA) did not give a synergistic effect on c-fos expression. Pretreatment of cells with PMA had no effect on [3H]PAF binding, but abolished the PAF-induced gene expression. PAF-stimulated gene expression was desensitized if cells were pretreated with PAF. Interestingly, epidermal growth factor was able to stimulate c-fos expression in PAF-desensitized cells, and thus indicated involvement of distinct mechanisms for the two stimuli. Forsholin, an activator of adenylate cyclase, did not induce c-fos expression and had no effect on the PAF response. Exposure of cells to PAF for as little as 1 min, followed by its removal, was sufficient to activate the gene expression and demonstrated the rapidity and the exquisite nature of the signalling involved in this process. It is concluded that activation of PAF receptor (a proposed G-protein-coupled receptor) causes rapid production of signals which induce the expression of c-fos gene and that this is mediated via tyrosine kinase and protein kinase C.

Original languageEnglish (US)
Pages (from-to)527-533
Number of pages7
JournalBiochemical Journal
Volume286
Issue number2
DOIs
StatePublished - Jan 1 1992

Fingerprint

fos Genes
Platelet Activating Factor
Gene expression
Protein-Tyrosine Kinases
Protein Kinase C
Gene Expression
Staurosporine
Genistein
protein kinase C kinase
Acetates
Genes
Cells
Protein C Inhibitor
Cycloheximide
Protein Kinase Inhibitors
G-Protein-Coupled Receptors
Adenylyl Cyclases
Epidermal Growth Factor
Protein Kinases
Inhibitory Concentration 50

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Involvement of tyrosine kinase and protein kinase C in platelet-activating-factor-induced c-fos gene expression in A-431 cells. / Tripathi, Y. B.; Lim, R. W.; Fernandez-Gallardo, S.; Kandala, J. C.; Guntaka, Ramareddy; Shukla, S. D.

In: Biochemical Journal, Vol. 286, No. 2, 01.01.1992, p. 527-533.

Research output: Contribution to journalArticle

Tripathi, Y. B. ; Lim, R. W. ; Fernandez-Gallardo, S. ; Kandala, J. C. ; Guntaka, Ramareddy ; Shukla, S. D. / Involvement of tyrosine kinase and protein kinase C in platelet-activating-factor-induced c-fos gene expression in A-431 cells. In: Biochemical Journal. 1992 ; Vol. 286, No. 2. pp. 527-533.
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