Protein kinase B/Akt binds and phosphorylates PED/PEA-15, stabilizing its antiapoptotic action

Alessandra Trencia, Anna Perfetti, Angela Cassese, Giovanni Vigliotta, Claudia Miele, Francesco Oriente, Stefania Santopietro, Ferdinando Giacco, Gerolama Condorelli, Pietro Formisano, Francesco Beguinot

Research output: Contribution to journalArticle

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Abstract

The antiapoptotic protein PED/PEA-15 features an Akt phosphorylation motif upstream from Ser116. In vitro, recombinant PED/PEA-15 was phosphorylated by Akt with a stoichiometry close to 1. Based on Western blotting with specific phospho-Ser116 PED/PEA-15 antibodies, Akt phosphorylation of PED/PEA-15 occurred mainly at Ser116. In addition, a mutant of PED/PEA-15 featuring the substitution of Ser116→Gly (PEDS116→G) showed 10-fold-decreased phosphorylation by Akt. In intact 293 cells, Akt also induced phosphorylation of PED/PEA-15 at Ser116. Based on pull-down and coprecipitation assays, PED/PEA-15 specifically bound Akt, independently of Akt activity. Serum activation of Akt as well as BAD phosphorylation by Akt showed no difference in 293 cells transfected with PED/PEA-15 and in untransfected cells (which express no endogenous PED/PEA-15). However, the antiapoptotic action of PED/PEA-15 was almost twofold reduced in PED116→G compared to that in PED/PEA-15WT cells. PED/PEA-15 stability closely paralleled Akt activation by serum in 293 cells. In these cells, the nonphosphorylatable PEDS116→G mutant exhibited a degradation rate threefold greater than that observed with wild-type PED/PEA-15. In the U373MG glioma cells, blocking Akt also reduced PED/PEA-15 levels and induced sensitivity to tumor necrosis factor-related apoptosis-inducing ligand apoptosis. Thus, phosphorylation by Akt regulates the antiapoptotic function of PED/PEA-15 at least in part by controlling the stability of PED/PEA-15. In part, Akt survival signaling may be mediated by PED/PEA-15.

Original languageEnglish (US)
Pages (from-to)4511-4521
Number of pages11
JournalMolecular and Cellular Biology
Volume23
Issue number13
DOIs
StatePublished - Jul 2003
Externally publishedYes

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Proto-Oncogene Proteins c-akt
Phosphorylation
Apoptosis
Serum
Glioma
Tumor Necrosis Factor-alpha
Western Blotting
Ligands
Antibodies

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Trencia, A., Perfetti, A., Cassese, A., Vigliotta, G., Miele, C., Oriente, F., ... Beguinot, F. (2003). Protein kinase B/Akt binds and phosphorylates PED/PEA-15, stabilizing its antiapoptotic action. Molecular and Cellular Biology, 23(13), 4511-4521. https://doi.org/10.1128/MCB.23.13.4511-4521.2003

Protein kinase B/Akt binds and phosphorylates PED/PEA-15, stabilizing its antiapoptotic action. / Trencia, Alessandra; Perfetti, Anna; Cassese, Angela; Vigliotta, Giovanni; Miele, Claudia; Oriente, Francesco; Santopietro, Stefania; Giacco, Ferdinando; Condorelli, Gerolama; Formisano, Pietro; Beguinot, Francesco.

In: Molecular and Cellular Biology, Vol. 23, No. 13, 07.2003, p. 4511-4521.

Research output: Contribution to journalArticle

Trencia, A, Perfetti, A, Cassese, A, Vigliotta, G, Miele, C, Oriente, F, Santopietro, S, Giacco, F, Condorelli, G, Formisano, P & Beguinot, F 2003, 'Protein kinase B/Akt binds and phosphorylates PED/PEA-15, stabilizing its antiapoptotic action', Molecular and Cellular Biology, vol. 23, no. 13, pp. 4511-4521. https://doi.org/10.1128/MCB.23.13.4511-4521.2003
Trencia, Alessandra ; Perfetti, Anna ; Cassese, Angela ; Vigliotta, Giovanni ; Miele, Claudia ; Oriente, Francesco ; Santopietro, Stefania ; Giacco, Ferdinando ; Condorelli, Gerolama ; Formisano, Pietro ; Beguinot, Francesco. / Protein kinase B/Akt binds and phosphorylates PED/PEA-15, stabilizing its antiapoptotic action. In: Molecular and Cellular Biology. 2003 ; Vol. 23, No. 13. pp. 4511-4521.
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