TMEM166/EVA1A interacts with ATG16L1 and induces autophagosome formation and cell death

Cell Death Dis. 2016 Aug 4;7(8):e2323. doi: 10.1038/cddis.2016.230.


The formation of the autophagosome is controlled by an orderly action of ATG proteins. However, how these proteins are recruited to autophagic membranes remain poorly clarified. In this study, we have provided a line of evidence confirming that EVA1A (eva-1 homolog A)/TMEM166 (transmembrane protein 166) is associated with autophagosomal membrane development. This notion is based on dotted EVA1A structures that colocalize with ZFYVE1, ATG9, LC3B, ATG16L1, ATG5, STX17, RAB7 and LAMP1, which represent different stages of the autophagic process. It is required for autophagosome formation as this phenotype was significantly decreased in EVA1A-silenced cells and Eva1a KO MEFs. EVA1A-induced autophagy is independent of the BECN1-PIK3C3 (phosphatidylinositol 3-kinase, catalytic subunit type 3) complex but requires ATG7 activity and the ATG12-ATG5/ATG16L1 complex. Here, we present a molecular mechanism by which EVA1A interacts with the WD repeats of ATG16L1 through its C-terminal and promotes ATG12-ATG5/ATG16L1 complex recruitment to the autophagic membrane and enhances the formation of the autophagosome. We also found that both autophagic and apoptotic mechanisms contributed to EVA1A-induced cell death while inhibition of autophagy and apoptosis attenuated EVA1A-induced cell death. Overall, these findings provide a comprehensive view to our understanding of the pathways involved in the role of EVA1A in autophagy and programmed cell death.

MeSH terms

  • Animals
  • Apoptosis*
  • Autophagosomes / metabolism*
  • Autophagy
  • Autophagy-Related Protein 5 / metabolism
  • Autophagy-Related Proteins / metabolism*
  • Beclin-1
  • Cell Line, Tumor
  • Gene Knockdown Techniques
  • Humans
  • Intracellular Membranes / metabolism
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism*
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Mutant Proteins / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism
  • Protein Binding
  • Structure-Activity Relationship


  • ATG16L1 protein, human
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Beclin-1
  • Becn1 protein, mouse
  • EVA1A protein, human
  • Map1lc3b protein, mouse
  • Membrane Proteins
  • Microtubule-Associated Proteins
  • Mutant Proteins