LRRK2 controls an EndoA phosphorylation cycle in synaptic endocytosis.
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.
Pubmed ID: 22998870 RIS Download
Acyltransferases | Animals | Animals, Genetically Modified | Brain | CHO Cells | Calcium | Clathrin | Cricetinae | Drosophila | Drosophila Proteins | Endocytosis | Gene Expression Regulation | Green Fluorescent Proteins | Humans | Mass Spectrometry | Mice | Microscopy, Electron, Transmission | Models, Molecular | Mutation | Neuromuscular Junction | Phosphorylation | Protein-Serine-Threonine Kinases | RNA, Small Interfering | Sequence Alignment | Serine | Synaptic Potentials | Synaptic Vesicles | Transfection