TBK1-mediated phosphorylation of LC3C and GABARAP-L2 controls autophagosome shedding by ATG4 protease

EMBO Rep. 2020 Jan 7;21(1):e48317. doi: 10.15252/embr.201948317. Epub 2019 Nov 11.


Autophagy is a highly conserved catabolic process through which defective or otherwise harmful cellular components are targeted for degradation via the lysosomal route. Regulatory pathways, involving post-translational modifications such as phosphorylation, play a critical role in controlling this tightly orchestrated process. Here, we demonstrate that TBK1 regulates autophagy by phosphorylating autophagy modifiers LC3C and GABARAP-L2 on surface-exposed serine residues (LC3C S93 and S96; GABARAP-L2 S87 and S88). This phosphorylation event impedes their binding to the processing enzyme ATG4 by destabilizing the complex. Phosphorylated LC3C/GABARAP-L2 cannot be removed from liposomes by ATG4 and are thus protected from ATG4-mediated premature removal from nascent autophagosomes. This ensures a steady coat of lipidated LC3C/GABARAP-L2 throughout the early steps in autophagosome formation and aids in maintaining a unidirectional flow of the autophagosome to the lysosome. Taken together, we present a new regulatory mechanism of autophagy, which influences the conjugation and de-conjugation of LC3C and GABARAP-L2 to autophagosomes by TBK1-mediated phosphorylation.

Keywords: ATG4; ATG8; TBK1; autophagy; phosphorylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autophagosomes* / metabolism
  • Autophagy
  • Autophagy-Related Protein 8 Family / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Microtubule-Associated Proteins* / genetics
  • Microtubule-Associated Proteins* / metabolism
  • Peptide Hydrolases
  • Phosphorylation


  • Autophagy-Related Protein 8 Family
  • Microtubule-Associated Proteins
  • Peptide Hydrolases