Skp2-Mediated RagA Ubiquitination Elicits a Negative Feedback to Prevent Amino-Acid-Dependent mTORC1 Hyperactivation by Recruiting GATOR1

Mol Cell. 2015 Jun 18;58(6):989-1000. doi: 10.1016/j.molcel.2015.05.010. Epub 2015 Jun 4.

Abstract

The regulation of RagA(GTP) is important for amino-acid-induced mTORC1 activation. Although GATOR1 complex has been identified as a negative regulator for mTORC1 by hydrolyzing RagA(GTP), how GATOR1 is recruited to RagA to attenuate mTORC1 signaling remains unclear. Moreover, how mTORC1 signaling is terminated upon amino acid stimulation is also unknown. We show that the recruitment of GATOR1 to RagA is induced by amino acids in an mTORC1-dependent manner. Skp2 E3 ligase drives K63-linked ubiquitination of RagA, which facilitates GATOR1 recruitment and RagA(GTP) hydrolysis, thereby providing a negative feedback loop to attenuate mTORC1 lysosomal recruitment and prevent mTORC1 hyperactivation. We further demonstrate that Skp2 promotes autophagy but inhibits cell size and cilia growth through RagA ubiquitination and mTORC1 inhibition. We thereby propose a negative feedback whereby Skp2-mediated RagA ubiquitination recruits GATOR1 to restrict mTORC1 signaling upon sustained amino acid stimulation, which serves a critical mechanism to maintain proper cellular functions.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / pharmacology*
  • Animals
  • Autophagy / genetics
  • Cell Line, Tumor
  • Enzyme Activation / drug effects
  • Feedback, Physiological / drug effects
  • Guanosine Triphosphate / metabolism
  • HEK293 Cells
  • Humans
  • Immunoblotting
  • Lysine / metabolism
  • Lysosomes / metabolism
  • Mechanistic Target of Rapamycin Complex 1
  • Mice
  • Mice, Knockout
  • Microscopy, Confocal
  • Models, Biological
  • Monomeric GTP-Binding Proteins / metabolism*
  • Multiprotein Complexes / metabolism*
  • NIH 3T3 Cells
  • Protein Binding / drug effects
  • RNA Interference
  • S-Phase Kinase-Associated Proteins / genetics
  • S-Phase Kinase-Associated Proteins / metabolism*
  • TOR Serine-Threonine Kinases / metabolism*
  • Ubiquitination / drug effects

Substances

  • Amino Acids
  • Multiprotein Complexes
  • S-Phase Kinase-Associated Proteins
  • Guanosine Triphosphate
  • Mechanistic Target of Rapamycin Complex 1
  • TOR Serine-Threonine Kinases
  • RRAGA protein, human
  • Monomeric GTP-Binding Proteins
  • Lysine