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.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acyltransferases / genetics
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Acyltransferases / metabolism*
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Animals
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Animals, Genetically Modified
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Brain / cytology
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Brain / metabolism
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CHO Cells
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Calcium / metabolism
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Clathrin / metabolism
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Cricetinae
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Drosophila
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Drosophila Proteins / genetics
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Drosophila Proteins / metabolism*
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Endocytosis / genetics
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Endocytosis / physiology*
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Gene Expression Regulation / genetics
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Humans
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Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
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Mass Spectrometry
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Mice
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Microscopy, Electron, Transmission
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Models, Molecular
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Mutation / genetics
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Neuromuscular Junction / drug effects
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Neuromuscular Junction / physiology*
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Neuromuscular Junction / ultrastructure
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Phosphorylation / genetics
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / metabolism*
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Sequence Alignment
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Serine / genetics
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Serine / metabolism
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Synaptic Potentials / drug effects
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Synaptic Potentials / genetics
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Synaptic Vesicles / drug effects
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Synaptic Vesicles / physiology
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Transfection
Substances
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Clathrin
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Drosophila Proteins
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RNA, Small Interfering
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Green Fluorescent Proteins
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Serine
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Acyltransferases
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endophilin A, Drosophila
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LRRK2 protein, human
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Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
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Protein Serine-Threonine Kinases
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Calcium