The Gβγ-Src signaling pathway regulates TNF-induced necroptosis via control of necrosome translocation

Cell Res. 2014 Apr;24(4):417-32. doi: 10.1038/cr.2014.17. Epub 2014 Feb 11.

Abstract

Formation of multi-component signaling complex necrosomes is essential for tumor necrosis factor α (TNF)-induced programmed necrosis (also called necroptosis). However, the mechanisms of necroptosis are still largely unknown. We isolated a TNF-resistant L929 mutant cell line generated by retrovirus insertion and identified that disruption of the guanine nucleotide-binding protein γ 10 (Gγ10) gene is responsible for this phenotype. We further show that Gγ10 is involved in TNF-induced necroptosis and Gβ2 is the partner of Gγ10. Src is the downstream effector of Gβ2γ10 in TNF-induced necroptosis because TNF-induced Src activation was impaired upon Gγ10 knockdown. Gγ10 does not affect TNF-induced activation of NF-κB and MAPKs and the formation of necrosomes, but is required for trafficking of necrosomes to their potential functioning site, an unidentified subcellular organelle that can be fractionated into heterotypic membrane fractions. The TNF-induced Gβγ-Src signaling pathway is independent of RIP1/RIP3 kinase activity and necrosome formation, but is required for the necrosome to function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Apoptosis / drug effects*
  • Apoptosis / genetics*
  • Base Sequence
  • Biological Transport / drug effects
  • Biological Transport / genetics
  • Cytoplasmic Vesicles / drug effects*
  • Cytoplasmic Vesicles / metabolism
  • GTP-Binding Protein beta Subunits / physiology*
  • GTP-Binding Protein gamma Subunits / physiology*
  • HEK293 Cells
  • Humans
  • Mice
  • Molecular Sequence Data
  • Necrosis / chemically induced
  • Necrosis / genetics
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • Tumor Cells, Cultured
  • Tumor Necrosis Factor-alpha / pharmacology*
  • src-Family Kinases / physiology*

Substances

  • GTP-Binding Protein beta Subunits
  • GTP-Binding Protein gamma Subunits
  • Tumor Necrosis Factor-alpha
  • src-Family Kinases