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Dissecting the effects of GTPase and kinase domain mutations on LRRK2 endosomal localization and activity

Capria Rinaldi1, Christopher S Waters1, Zizheng Li1

  • 1Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA.

Cell Reports
|May 4, 2023
PubMed

Insights

Mutant leucine-rich repeat kinase 2 (LRRK2) in Parkinson's disease localizes differently, with GTPase-inactivating mutations forming more endosomes and increasing Rab phosphorylation compared to kinase-activating mutations.

Area of Science:

  • Cell Biology
  • Neuroscience
  • Biochemistry

Background:

  • Mutations in leucine-rich repeat kinase 2 (LRRK2) are a major genetic cause of Parkinson's disease.
  • LRRK2 mutations are associated with altered Rab GTPase phosphorylation, but the mechanism is unclear.
  • GTPase-inactivating LRRK2 mutations paradoxically increase Rab phosphorylation more than kinase-activating mutations.

Purpose of the Study:

  • To investigate if differential cellular localization of mutant LRRK2 explains varying Rab phosphorylation levels.
  • To explore the role of endosomal maturation and positive feedback in LRRK2 localization and activity.

Main Methods:

  • Utilized cell-based assays to observe LRRK2 localization and Rab phosphorylation.
  • Manipulated endosomal maturation to induce LRRK2+ endosome formation.
  • Analyzed localization patterns of various LRRK2 mutants.

Main Results:

  • Blocking endosomal maturation rapidly formed mutant LRRK2+ endosomes, sites of Rab phosphorylation.
  • LRRK2+ endosomes exhibited positive feedback, reinforcing LRRK2 and pRab localization.
  • GTPase-inactivating LRRK2 mutants formed significantly more LRRK2+ endosomes than kinase-activating mutants.
  • This resulted in higher overall cellular levels of phosphorylated Rab GTPases.

Conclusions:

  • Altered cellular localization, specifically increased retention on intracellular membranes, underlies the higher Rab phosphorylation observed with GTPase-inactivating LRRK2 mutants.
  • This finding provides a novel mechanistic link between LRRK2 mutation type, subcellular localization, and Parkinson's disease pathogenesis.
Keywords:
CP: Cell biologyLRRK2Parkinson's diseaseRab phosphorylationVPS34 inhibitionendosomal localizationpositive feedback

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