MECHANISMS OF SIGNAL TRANSDUCTION
Endothelin-1 Induces Serine Phosphorylation of the Adaptor Protein p66Shc and Its Association with 14-3-3 Protein in Glomerular Mesangial Cells*

https://doi.org/10.1074/jbc.M102008200Get rights and content
Under a Creative Commons license
open access

Endothelin-1 (ET-1) is a vasoconstrictor peptide known to be a potent mitogen for glomerular mesangial cells (GMC). In the current study, it is demonstrated that ET-1 treatment of GMC results in serine phosphorylation of the 66-kDa isoform of the adapter protein Shc (p66Shc). ET-1-induced serine phosphorylation of p66Shc requires activation of the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling module and is efficiently inhibited by both a MAPK/ERK kinase (MEK)-selective inhibitor and adenovirus-mediated transfer of a dominant interfering MEK1 mutant. Furthermore, adenovirus-mediated transfer of a constitutively active MEK1 mutant was found to markedly increase p66Shc serine phosphorylation. Adenoviruses encoding constitutively active mutants of MAPK kinases 3 and 6 (upstream kinases of p38MAPK) and 7 (upstream kinase of c-Jun NH2-terminal kinase) failed to induce serine phosphorylation of this adaptor protein. Serine phosphorylation of p66Shc resulted in its association with the serine binding motif-containing protein 14-3-3. ET-1-induced phosphorylation of a serine encompassed in the 14-3-3 binding motif of p66Shcwas confirmed in experiments employing anti-phospho-14-3-3 binding motif antibodies. These studies are the first to demonstrate that G protein-coupled receptors stimulate serine phosphorylation of p66Shc and the first to report the formation of a signaling complex between p66Shc and 14-3-3.

Cited by (0)

Published, JBC Papers in Press, May 7, 2001, DOI 10.1074/jbc.M102008200

*

This work was supported by a grant from Ministero dell'Universita e della Ricerca Scientifica e Tecnologica (MURST) and the University of Florence and by National Institutes of Health Research Grants DK 41684 and HL 22563.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§

Correspondence may be addressed. Tel.: 39-055-4296479; Fax: 39-055-417123; E-mail: [email protected].

**

To whom correspondence may be addressed: Tel.: 414-456-4438; Fax: 414-456-6515; E-mail: [email protected].