K27-linked ubiquitination of BRAF by ITCH engages cytokine response to maintain MEK-ERK signaling

Nat Commun. 2019 Apr 23;10(1):1870. doi: 10.1038/s41467-019-09844-0.

Abstract

BRAF plays an indispensable role in activating the MEK/ERK pathway to drive tumorigenesis. Receptor tyrosine kinase and RAS-mediated BRAF activation have been extensively characterized, however, it remains undefined how BRAF function is fine-tuned by stimuli other than growth factors. Here, we report that in response to proinflammatory cytokines, BRAF is subjected to lysine 27-linked poly-ubiquitination in melanoma cells by the ITCH ubiquitin E3 ligase. Lysine 27-linked ubiquitination of BRAF recruits PP2A to antagonize the S365 phosphorylation and disrupts the inhibitory interaction with 14-3-3, leading to sustained BRAF activation and subsequent elevation of the MEK/ERK signaling. Physiologically, proinflammatory cytokines activate ITCH to maintain BRAF activity and to promote proliferation and invasion of melanoma cells, whereas the ubiquitination-deficient BRAF mutant displays compromised kinase activity and reduced tumorigenicity. Collectively, our study reveals a pivotal role for ITCH-mediated BRAF ubiquitination in coordinating the signals between cytokines and the MAPK pathway activation in melanoma cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 14-3-3 Proteins / metabolism
  • Animals
  • Cell Line, Tumor
  • Cell Survival
  • Cell Transformation, Neoplastic*
  • Cytokines / metabolism*
  • HEK293 Cells
  • Humans
  • Inflammation Mediators / metabolism
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • Lysine / metabolism
  • MAP Kinase Signaling System*
  • Male
  • Melanoma / pathology
  • Mice
  • Mice, Nude
  • Phosphorylation
  • Proto-Oncogene Proteins B-raf / metabolism*
  • Repressor Proteins / metabolism*
  • Skin Neoplasms / pathology
  • Ubiquitin-Protein Ligases / metabolism*
  • Ubiquitination
  • Xenograft Model Antitumor Assays

Substances

  • 14-3-3 Proteins
  • Cytokines
  • Inflammation Mediators
  • Repressor Proteins
  • ITCH protein, human
  • Ubiquitin-Protein Ligases
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • JNK Mitogen-Activated Protein Kinases
  • Lysine