The SAC1 domain in synaptojanin is required for autophagosome maturation at presynaptic terminals

EMBO J. 2017 May 15;36(10):1392-1411. doi: 10.15252/embj.201695773. Epub 2017 Mar 22.

Abstract

Presynaptic terminals are metabolically active and accrue damage through continuous vesicle cycling. How synapses locally regulate protein homeostasis is poorly understood. We show that the presynaptic lipid phosphatase synaptojanin is required for macroautophagy, and this role is inhibited by the Parkinson's disease mutation R258Q. Synaptojanin drives synaptic endocytosis by dephosphorylating PI(4,5)P2, but this function appears normal in SynaptojaninRQ knock-in flies. Instead, R258Q affects the synaptojanin SAC1 domain that dephosphorylates PI(3)P and PI(3,5)P2, two lipids found in autophagosomal membranes. Using advanced imaging, we show that SynaptojaninRQ mutants accumulate the PI(3)P/PI(3,5)P2-binding protein Atg18a on nascent synaptic autophagosomes, blocking autophagosome maturation at fly synapses and in neurites of human patient induced pluripotent stem cell-derived neurons. Additionally, we observe neurodegeneration, including dopaminergic neuron loss, in SynaptojaninRQ flies. Thus, synaptojanin is essential for macroautophagy within presynaptic terminals, coupling protein turnover with synaptic vesicle cycling and linking presynaptic-specific autophagy defects to Parkinson's disease.

Keywords: Parkinson's disease; correlative light and electron microscopy; induced pluripotent stem cells; single‐molecule tracking; synapse.

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Autophagosomes / metabolism*
  • Autophagy*
  • Autophagy-Related Proteins / analysis
  • Cells, Cultured
  • Drosophila
  • Humans
  • Membrane Proteins / analysis
  • Mutation, Missense
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Parkinson Disease / pathology
  • Phosphatidylinositol Phosphates / metabolism
  • Phosphoric Monoester Hydrolases / genetics
  • Phosphoric Monoester Hydrolases / metabolism*
  • Presynaptic Terminals / enzymology*
  • Presynaptic Terminals / metabolism*

Substances

  • Autophagy-Related Proteins
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Phosphatidylinositol Phosphates
  • WIPI1 protein, human
  • phosphatidylinositol 3,5-diphosphate
  • phosphatidylinositol 3-phosphate
  • synaptojanin
  • Phosphoric Monoester Hydrolases