Phosphorylation site dynamics of early T-cell receptor signaling

PLoS One. 2014 Aug 22;9(8):e104240. doi: 10.1371/journal.pone.0104240. eCollection 2014.

Abstract

In adaptive immune responses, T-cell receptor (TCR) signaling impacts multiple cellular processes and results in T-cell differentiation, proliferation, and cytokine production. Although individual protein-protein interactions and phosphorylation events have been studied extensively, we lack a systems-level understanding of how these components cooperate to control signaling dynamics, especially during the crucial first seconds of stimulation. Here, we used quantitative proteomics to characterize reshaping of the T-cell phosphoproteome in response to TCR/CD28 co-stimulation, and found that diverse dynamic patterns emerge within seconds. We detected phosphorylation dynamics as early as 5 s and observed widespread regulation of key TCR signaling proteins by 30 s. Development of a computational model pointed to the presence of novel regulatory mechanisms controlling phosphorylation of sites with central roles in TCR signaling. The model was used to generate predictions suggesting unexpected roles for the phosphatase PTPN6 (SHP-1) and shortcut recruitment of the actin regulator WAS. Predictions were validated experimentally. This integration of proteomics and modeling illustrates a novel, generalizable framework for solidifying quantitative understanding of a signaling network and for elucidating missing links.

Publication types

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

MeSH terms

  • Cell Line
  • Humans
  • Models, Biological
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Protein Interaction Maps
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6 / metabolism
  • Proteome
  • Proteomics / methods
  • Receptors, Antigen, T-Cell / metabolism*
  • Signal Transduction*
  • T-Lymphocytes / metabolism*
  • Tyrosine / metabolism
  • Wiskott-Aldrich Syndrome Protein Family / metabolism

Substances

  • Phosphoproteins
  • Proteome
  • Receptors, Antigen, T-Cell
  • Wiskott-Aldrich Syndrome Protein Family
  • Tyrosine
  • PTPN6 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 6