Phosphoproteomics reveals resveratrol-dependent inhibition of Akt/mTORC1/S6K1 signaling

J Proteome Res. 2014 Dec 5;13(12):5734-42. doi: 10.1021/pr500714a. Epub 2014 Oct 29.

Abstract

Resveratrol, a plant-derived polyphenol, regulates many cellular processes, including cell proliferation, aging and autophagy. However, the molecular mechanisms of resveratrol action in cells are not completely understood. Intriguingly, resveratrol treatment of cells growing in nutrient-rich conditions induces autophagy, while acute resveratrol treatment of cells in a serum-deprived state inhibits autophagy. In this study, we performed a phosphoproteomic analysis after applying resveratrol to serum-starved cells with the goal of identifying the acute signaling events initiated by resveratrol in a serum-deprived state. We determined that resveratrol in serum-starved conditions reduces the phosphorylation of several proteins belonging to the mTORC1 signaling pathway, most significantly, PRAS40 at T246 and S183. Under these same conditions, we also found that resveratrol altered the phosphorylation of several proteins involved in various biological processes, most notably transcriptional modulators, represented by p53, FOXA1, and AATF. Together these data provide a more comprehensive view of both the spectrum of phosphoproteins upon which resveratrol acts as well as the potential mechanisms by which it inhibits autophagy in serum-deprived cells.

Keywords: PRAS40; S6K1; SILAC; mTORC1; phosphoproteomics; resveratrol.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology
  • Autophagy / drug effects
  • Chromatography, Liquid
  • Culture Media, Serum-Free / pharmacology
  • Humans
  • Immunoblotting
  • MCF-7 Cells
  • Mechanistic Target of Rapamycin Complex 1
  • Models, Biological
  • Multiprotein Complexes / metabolism*
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphopeptides / metabolism
  • Phosphoproteins / metabolism*
  • Phosphorylation / drug effects
  • Proteome / metabolism
  • Proteomics / methods*
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Resveratrol
  • Ribosomal Protein S6 Kinases, 70-kDa / metabolism*
  • Signal Transduction / drug effects
  • Stilbenes / pharmacology*
  • TOR Serine-Threonine Kinases / metabolism*
  • Tandem Mass Spectrometry

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Culture Media, Serum-Free
  • Multiprotein Complexes
  • Phosphopeptides
  • Phosphoproteins
  • Proteome
  • Stilbenes
  • AKT1 protein, human
  • Mechanistic Target of Rapamycin Complex 1
  • Proto-Oncogene Proteins c-akt
  • Ribosomal Protein S6 Kinases, 70-kDa
  • TOR Serine-Threonine Kinases
  • ribosomal protein S6 kinase, 70kD, polypeptide 1
  • Resveratrol