RAB3 phosphorylation by pathogenic LRRK2 impairs trafficking of synaptic vesicle precursors

Abstract

Gain-of-function mutations in the LRRK2 gene cause Parkinson’s disease (PD), characterized by debilitating motor and non-motor symptoms. Increased phosphorylation of a subset of RAB GTPases by LRRK2 is implicated in PD pathogenesis. We find that increased phosphorylation of RAB3A, a cardinal synaptic vesicle precursor (SVP) protein, disrupts anterograde axonal transport of SVPs in iPSC-derived human neurons (iNeurons) expressing hyperactive LRRK2-p.R1441H. Knockout of the opposing protein phosphatase 1H (PPM1H) in iNeurons phenocopies this effect. In these models, the compartmental distribution of synaptic proteins is altered; synaptophysin and synaptobrevin-2 become sequestered in the neuronal soma with decreased delivery to presynaptic sites along the axon. We find that RAB3A phosphorylation disrupts binding to the motor adaptor MADD, potentially preventing the formation of the RAB3A–MADD-KIF1A/1Bβ complex driving anterograde SVP transport. RAB3A hyperphosphorylation also disrupts interactions with RAB3GAP and RAB-GDI1. Our results reveal a mechanism by which pathogenic hyperactive LRRK2 may contribute to the altered synaptic homeostasis associated with characteristic non-motor and cognitive manifestations of PD.

Presynaptic perspective: Axonal transport defects in neurodevelopmental disorders

“…Thus, DENN/MADD connects kinesin-3 with SVPs carrying GTP-bound Rab3. RAB3A phosphorylation disrupts its binding to MADD, thus preventing SVP loading onto KIF1A/KIF1Bβ motors for anterograde transport ( Dou et al, 2024 ). In DENN/MADD knockout mice, SV numbers are reduced ( Tanaka et al, 2001 ).…”