Phosphoproteomic comparison of Pik3ca and Pten signalling identifies the nucleotidase NT5C as a novel AKT substrate

Sci Rep. 2017 Jan 6:7:39985. doi: 10.1038/srep39985.

Abstract

To identify novel effectors and processes regulated by PI3K pathway activation, we performed an unbiased phosphoproteomic screen comparing two common events of PI3K deregulation in cancer: oncogenic Pik3ca mutation (Pik3caH1047R) and deletion of Pten. Using mouse embryonic fibroblast (MEF) models that generate inducible, low-level pathway activation as observed in cancer, we quantified 7566 unique phosphopeptides from 3279 proteins. A number of proteins were found to be differentially-regulated by Pik3caH1047R and Pten loss, suggesting unique roles for these two events in processes such as vesicular trafficking, DNA damage repair and RNA splicing. We also identified novel PI3K effectors that were commonly-regulated, including putative AKT substrates. Validation of one of these hits, confirmed NT5C (5',3'-Nucleotidase, Cytosolic) as a novel AKT substrate, with an unexpected role in actin cytoskeleton regulation via an interaction with the ARP2/3 complex. This study has produced a comprehensive data resource and identified a new link between PI3K pathway activation and actin regulation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5'-Nucleotidase / metabolism*
  • Animals
  • Cells, Cultured
  • Class I Phosphatidylinositol 3-Kinases
  • Gene Deletion
  • Gene Expression Regulation
  • Mice
  • Mouse Embryonic Stem Cells
  • Mutation
  • PTEN Phosphohydrolase / genetics*
  • PTEN Phosphohydrolase / metabolism
  • Phosphatidylinositol 3-Kinases / genetics*
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphoproteins / analysis*
  • Proteomics / methods*
  • Proto-Oncogene Proteins c-akt / metabolism*
  • Signal Transduction

Substances

  • Phosphoproteins
  • Class I Phosphatidylinositol 3-Kinases
  • Pik3ca protein, mouse
  • Proto-Oncogene Proteins c-akt
  • 5'-Nucleotidase
  • PTEN Phosphohydrolase
  • Pten protein, mouse