Phosphorylation of p90 and p52 in response to phorbol-esters in Swiss/3T3 cells overexpressing protein kinase C-α

Hagit Eldar, Etta Livneh

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Cell lines stably overexpressing protein kinase C (PKC)-α were previously described by us. These cell lines were generated by the introduction of the full length cDNA coding for PKC-α into Swiss/3T3 cells. Here we show that activation of PKC-α by phorbol-esters induced in these cells specific phosphorylation of two cellular proteins p90 and p52. Phosphorylation of p80 (MARCKS protein), previously identified as a substrate for PKC, was also enhanced. Phosphorylated p90 and p52 proteins were associated with particulate membrane-enriched fractions and were extractable with the use of nonionic detergents. Time course analysis of phorbol-ester induced phosphorylation of p90 and p52 revealed maximal stimulation of phosphorylation after 15-30 min. Phosphamino acid analysis showed that phosphorylation of p90 and p52 occurred mainly on serine residues. Phosphorylation of p52 was also on threonine residues. Whereas, phorbol ester activation induced phosphorylation of both p90 and p52, the mitogens platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF) enhanced phosphorylation of p90, but not p52. Thus, our studies showed the involvement of PKC-α in the regulation of p90 and p52 phosphorylation and provided direct evidence for the role of PKC-α in cellular signaling by PDGF and FGF. Moreover, the fact that phosphorylation of p52 was specific to phorbol ester activation may suggest its involvement in tumor promotion. Characterization of p90 and p52 will enable us to reveal the phosphorylation cascade activated downstream to PKC-α and to determine their role in mitogenic signaling and tumor promotion.

Original languageEnglish
Pages (from-to)1049-1056
Number of pages8
JournalMolecular Biology of the Cell
Volume3
Issue number9
DOIs
StatePublished - 1 Jan 1992
Externally publishedYes

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