LRRK2 controls an EndoA phosphorylation cycle in synaptic endocytosis

Neuron. 2012 Sep 20;75(6):1008-21. doi: 10.1016/j.neuron.2012.08.022.

Abstract

LRRK2 is a kinase mutated in Parkinson's disease, but how the protein affects synaptic function remains enigmatic. We identified LRRK2 as a critical regulator of EndophilinA. Using genetic and biochemical studies involving Lrrk loss-of-function mutants and Parkinson-related LRRK2(G2019S) gain-of-kinase function, we show that LRRK2 affects synaptic endocytosis by phosphorylating EndoA at S75, a residue in the BAR domain. We show that LRRK2-mediated EndoA phosphorylation has profound effects on EndoA-dependent membrane tubulation and membrane association in vitro and in vivo and on synaptic vesicle endocytosis at Drosophila neuromuscular junctions in vivo. Our work uncovers a regulatory mechanism that indicates that reduced LRRK2 kinase activity facilitates EndoA membrane association, while increased kinase activity inhibits membrane association. Consequently, both too much and too little LRRK2-dependent EndoA phosphorylation impedes synaptic endocytosis, and we propose a model in which LRRK2 kinase activity is part of an EndoA phosphorylation cycle that facilitates efficient vesicle formation at synapses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyltransferases / genetics
  • Acyltransferases / metabolism*
  • Animals
  • Animals, Genetically Modified
  • Brain / cytology
  • Brain / metabolism
  • CHO Cells
  • Calcium / metabolism
  • Clathrin / metabolism
  • Cricetinae
  • Drosophila
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism*
  • Endocytosis / genetics
  • Endocytosis / physiology*
  • Gene Expression Regulation / genetics
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
  • Mass Spectrometry
  • Mice
  • Microscopy, Electron, Transmission
  • Models, Molecular
  • Mutation / genetics
  • Neuromuscular Junction / drug effects
  • Neuromuscular Junction / physiology*
  • Neuromuscular Junction / ultrastructure
  • Phosphorylation / genetics
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Sequence Alignment
  • Serine / genetics
  • Serine / metabolism
  • Synaptic Potentials / drug effects
  • Synaptic Potentials / genetics
  • Synaptic Vesicles / drug effects
  • Synaptic Vesicles / physiology
  • Transfection

Substances

  • Clathrin
  • Drosophila Proteins
  • RNA, Small Interfering
  • Green Fluorescent Proteins
  • Serine
  • Acyltransferases
  • endophilin A, Drosophila
  • LRRK2 protein, human
  • Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
  • Protein-Serine-Threonine Kinases
  • Calcium