PKA phosphorylation of Src mediates Rap1 activation in NGF and cAMP signaling in PC12 cells

J Cell Sci. 2004 Dec 1;117(Pt 25):6085-94. doi: 10.1242/jcs.01527. Epub 2004 Nov 16.

Abstract

Recent studies suggest that the tyrosine kinase Src plays an important role in the hormonal regulation of extracellular signal-regulated kinases (ERKs) via cyclic AMP (cAMP). Src has also been proposed to mediate signals downstream of nerve growth factor (NGF). Here, we report that the cAMP-dependent protein kinase A (PKA) induced the phosphorylation of Src at residue serine17 (S17) in multiple cell types including PC12, Hek293, AtT-20 and CHO cells. In PC12 cells, Src phosphorylation on S17 participates in the activation of the small G protein Rap1 by both cAMP and NGF. In these cells, Rap1 is required for cAMP/PKA signaling to ERKs and also for the sustained activation of ERKs by NGF. The activation of Rap1 by both cAMP and NGF was blocked by PP2, an inhibitor of Src family kinases, and by a Src mutant incapable of being phosphorylated by PKA (SrcS17A), consistent with the requirement of PKA phosphorylation of Src at S17 in these actions. PP2 and SrcS17A also inhibited the Rap1-dependent activation of ERKs by both agents. These results strongly indicate that PKA phosphorylation of Src at S17 is essential for cAMP and NGF signaling in PC12 cells and identify PKA as an important downstream target of NGF. PKA phosphorylation of Src may therefore be required for Rap1 activation in PC12 cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Western
  • CHO Cells
  • Cell Line
  • Cricetinae
  • Cyclic AMP / metabolism*
  • Cyclic AMP-Dependent Protein Kinases / metabolism*
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Humans
  • Immunoprecipitation
  • Nerve Growth Factor / metabolism*
  • Neurons / metabolism
  • PC12 Cells
  • Phosphorylation
  • Plasmids / metabolism
  • Rats
  • Serine / chemistry*
  • Signal Transduction
  • Time Factors
  • rap1 GTP-Binding Proteins / metabolism*
  • ras Proteins / metabolism
  • src-Family Kinases / metabolism*

Substances

  • Serine
  • Nerve Growth Factor
  • Cyclic AMP
  • src-Family Kinases
  • Cyclic AMP-Dependent Protein Kinases
  • Extracellular Signal-Regulated MAP Kinases
  • rap1 GTP-Binding Proteins
  • ras Proteins