The SAC1 domain in synaptojanin is required for autophagosome maturation at presynaptic terminals.
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 inSynaptojaninRQknock-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 thatSynaptojaninRQmutants 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, inSynaptojaninRQflies. 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.
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Roeland
R
Vanhauwaert
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Sabine
S
Kuenen
Department of Clinical Genetics, Erasmus MC, Rotterdam, The Netherlands.
Roy
R
Masius
Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Qld, Australia.
Adekunle
A
Bademosi
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Julia
J
Manetsberger
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Nils
N
Schoovaerts
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Laura
L
Bounti
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Serguei
S
Gontcharenko
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Jef
J
Swerts
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Sven
S
Vilain
Department of Medicine and Surgery, Center for Neurodegenerative Diseases (CEMAND), University of Salerno, Salerno, Italy.
Marina
M
Picillo
Department of Medicine and Surgery, Center for Neurodegenerative Diseases (CEMAND), University of Salerno, Salerno, Italy.
Paolo
P
Barone
Department of Psychiatry, Erasmus MC, Rotterdam, The Netherlands.
Shashini T
ST
Munshi
Department of Psychiatry, Erasmus MC, Rotterdam, The Netherlands.
Femke Ms
FM
de Vrij
Department of Psychiatry, Erasmus MC, Rotterdam, The Netherlands.
Steven A
SA
Kushner
VIB Center for Brain & Disease Research, Leuven, Belgium.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.|Electron Microscopy Platform, VIB Bio-Imaging Core, Leuven, Belgium.
Natalia V
NV
Gounko
Department of Clinical Genetics, Erasmus MC, Rotterdam, The Netherlands.
Wim
W
Mandemakers
Department of Clinical Genetics, Erasmus MC, Rotterdam, The Netherlands.
Vincenzo
V
Bonifati
Clem Jones Centre for Ageing Dementia Research, Queensland Brain Institute, The University of Queensland, Brisbane, Qld, Australia.
Frederic A
FA
Meunier
VIB Center for Brain & Disease Research, Leuven, Belgium sandra.soukup@cme.vib-kuleuven.be patrik.verstreken@cme.vib-kuleuven.be.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Sandra-Fausia
0000-0003-2915-919X
SF
Soukup
VIB Center for Brain & Disease Research, Leuven, Belgium sandra.soukup@cme.vib-kuleuven.be patrik.verstreken@cme.vib-kuleuven.be.|Department of Human Genetics, Leuven Institute for Neurodegenerative Disease (LIND), KU Leuven, Leuven, Belgium.
Patrik
0000-0002-5073-5393
P
Verstreken
D019943:Amino Acid Substitution; D000818:Animals; D000071182:Autophagosomes / Q000378:metabolism*; D001343:Autophagy; D000071183:Autophagy-Related Proteins / Q000032:analysis; D002478:Cells, Cultured; D004330:Drosophila; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D020125:Mutation, Missense; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010300:Parkinson Disease / Q000473:pathology; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D017729:Presynaptic Terminals / Q000201:enzymology* / Q000378:metabolism*
D000071183:Autophagy-Related Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D018129:Phosphatidylinositol Phosphates; C522812:WIPI1 protein, human; C106336:phosphatidylinositol 3,5-diphosphate; C055525:phosphatidylinositol 3-phosphate; C097702:synaptojanin; D010744:Phosphoric Monoester Hydrolases
Parkinson's disease; correlative light and electron microscopy; induced pluripotent stem cells; single‐molecule tracking; synapse