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

• DOI: 10.1038/cr.2014.17
• Publication Date: 2014-02-11T11:50:07Z
• Authors: Lisheng Li; Wanze Chen; Yaoji Liang; Huabin Ma; Wenjuan Li; Zhenru Zhou; Jie Li; Yan Ding; Junming Ren; Juan Lin; Felicia Han; Jianfeng Wu; Jiahuai Han
• Keywords: 0301 basic medicine; VOLTAGE-DEPENDENT MODULATION; 571; SMOOTH-MUSCLE-CELLS; Molecular Sequence Data; Apoptosis; Mice; Necrosis; 03 medical and health sciences; GTP-Binding Protein gamma Subunits; MIXED LINEAGE KINASE; Tumor Cells, Cultured; Animals; Humans; Amino Acid Sequence; PROGRAMMED NECROSIS; L929 CELLS; TYROSINE PHOSPHORYLATION; Base Sequence; Tumor Necrosis Factor-alpha; C-SRC; Cytoplasmic Vesicles; GTP-Binding Protein beta Subunits; Biological Transport; PROTEIN-COUPLED RECEPTOR; HEK293 Cells; src-Family Kinases; MEDIATED CYTOTOXICITY; INDUCED CELL-DEATH; Signal Transduction

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.