Global Identification of Small Ubiquitin-related Modifier (SUMO) Substrates Reveals Crosstalk between SUMOylation and Phosphorylation Promotes Cell Migration

Mol Cell Proteomics. 2018 May;17(5):871-888. doi: 10.1074/mcp.RA117.000014. Epub 2018 Feb 8.


Proteomics studies have revealed that SUMOylation is a widely used post-translational modification (PTM) in eukaryotes. However, how SUMO E1/2/3 complexes use different SUMO isoforms and recognize substrates remains largely unknown. Using a human proteome microarray-based activity screen, we identified over 2500 proteins that undergo SUMO E3-dependent SUMOylation. We next constructed a SUMO isoform- and E3 ligase-dependent enzyme-substrate relationship network. Protein kinases were significantly enriched among SUMOylation substrates, suggesting crosstalk between phosphorylation and SUMOylation. Cell-based analyses of tyrosine kinase, PYK2, revealed that SUMOylation at four lysine residues promoted PYK2 autophosphorylation at tyrosine 402, which in turn enhanced its interaction with SRC and full activation of the SRC-PYK2 complex. SUMOylation on WT but not the 4KR mutant of PYK2 further elevated phosphorylation of the downstream components in the focal adhesion pathway, such as paxillin and Erk1/2, leading to significantly enhanced cell migration during wound healing. These studies illustrate how our SUMO E3 ligase-substrate network can be used to explore crosstalk between SUMOylation and other PTMs in many biological processes.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Cell Movement*
  • HeLa Cells
  • Humans
  • Phosphorylation
  • Phosphotyrosine / metabolism
  • Protein Kinases / chemistry
  • Protein Kinases / metabolism
  • Proteomics
  • Reproducibility of Results
  • Signal Transduction
  • Small Ubiquitin-Related Modifier Proteins / metabolism*
  • Substrate Specificity
  • Sumoylation*
  • Ubiquitin-Protein Ligases / metabolism


  • Small Ubiquitin-Related Modifier Proteins
  • Phosphotyrosine
  • Ubiquitin-Protein Ligases
  • Protein Kinases