Characterization of early autophagy signaling by quantitative phosphoproteomics

Autophagy. 2014 Feb;10(2):356-71. doi: 10.4161/auto.26864. Epub 2013 Nov 21.


Under conditions of nutrient shortage autophagy is the primary cellular mechanism ensuring availability of substrates for continuous biosynthesis. Subjecting cells to starvation or rapamycin efficiently induces autophagy by inhibiting the MTOR signaling pathway triggering increased autophagic flux. To elucidate the regulation of early signaling events upon autophagy induction, we applied quantitative phosphoproteomics characterizing the temporal phosphorylation dynamics after starvation and rapamycin treatment. We obtained a comprehensive atlas of phosphorylation kinetics within the first 30 min upon induction of autophagy with both treatments affecting widely different cellular processes. The identification of dynamic phosphorylation already after 2 min demonstrates that the earliest events in autophagy signaling occur rapidly after induction. The data was subjected to extensive bioinformatics analysis revealing regulated phosphorylation sites on proteins involved in a wide range of cellular processes and an impact of the treatments on the kinome. To approach the potential function of the identified phosphorylation sites we performed a screen for MAP1LC3-interacting proteins and identified a group of binding partners exhibiting dynamic phosphorylation patterns. The data presented here provide a valuable resource on phosphorylation events underlying early autophagy induction.

Keywords: autophagy; bioinformatics; mass spectrometry; phosphoproteomics; phosphorylation; proteomics; signal transduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagy / drug effects*
  • Cell Line, Tumor
  • Humans
  • Phosphoproteins / metabolism
  • Phosphorylation / drug effects
  • Proteomics
  • Signal Transduction / drug effects*
  • Sirolimus / pharmacology*
  • Starvation / metabolism
  • Time Factors


  • Phosphoproteins
  • Sirolimus