Regulator of G-protein signaling 19 (RGS19) and its partner Gα-inhibiting activity polypeptide 3 (GNAI3) are required for zVAD-induced autophagy and cell death in L929 cells

PLoS One. 2014 Apr 21;9(4):e94634. doi: 10.1371/journal.pone.0094634. eCollection 2014.


Autophagy has diverse biological functions and is involved in many biological processes. The L929 cell death induced by the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-(OMe)-fluoromethyl ketone (zVAD) was shown to be an autophagy-mediated death for which RIP1 and RIP3 were both required. It was also reported that zVAD can induce a small amount of TNF production, which was shown to be required for zVAD-induced L929 cell death, arguing for the contribution of autophagy in the zVAD-induced L929 cell death. In an effort to study RIP3 mediated cell death, we identified regulator of G-protein signaling 19 (RGS19) as a RIP3 interacting protein. We showed that RGS19 and its partner Gα-inhibiting activity polypeptide 3 (GNAI3) are involved in zVAD-, but not TNF-, induced cell death. The role of RGS19 and GNAI3 in zVAD-induced cell death is that they are involved in zVAD-induced autophagy. By the use of small hairpin RNAs and chemical inhibitors, we further demonstrated that zVAD-induced autophagy requires not only RIP1, RIP3, PI3KC3 and Beclin-1, but also RGS19 and GNAI3, and this autophagy is required for zVAD-induced TNF production. Collectively, our data suggest that zVAD-induced L929 cell death is a synergistic result of autophagy, caspase inhibition and autocrine effect of TNF.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy / drug effects*
  • Cell Line, Tumor
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • GTP-Binding Protein alpha Subunits, Gi-Go / metabolism*
  • Gene Knockdown Techniques
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • MAP Kinase Signaling System / drug effects
  • Mice
  • Oligopeptides / pharmacology*
  • Protein Binding / drug effects
  • RGS Proteins / metabolism*
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism
  • Tumor Necrosis Factor-alpha / biosynthesis
  • Tumor Necrosis Factor-alpha / pharmacology


  • Oligopeptides
  • RGS Proteins
  • Tumor Necrosis Factor-alpha
  • benzyloxycarbonyl-valyl-alanyl-aspartic acid
  • regulator of G-protein signalling 19
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk3 protein, mouse
  • Extracellular Signal-Regulated MAP Kinases
  • JNK Mitogen-Activated Protein Kinases
  • GTP-Binding Protein alpha Subunits, Gi-Go
  • Gnai3 protein, mouse

Grant support

This work was supported by the Fundamental Research Funds for the Central Universities (2010121105), the Natural Science Foundation of Fujian Province of China (2011J05100), the National Basic Research Program of China (973 Program 2013CB944903, 2014CB541804), the National Natural Science Foundation of China (31330047, 91029304, 81061160512, 31221065), the Hi-Tech Research and Development Program of China (863 program; 2012AA02A201), the 111 Project (B12001), and the Open Research Fund of State Key Laboratory of Cellular Stress Biology, Xiamen University (SKLCSB2012KF003). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.