Related Experiment Video

Updated: Nov 12, 2025

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
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Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

Published on: December 14, 2017

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LRRK2-phosphorylated Rab10 sequesters Myosin Va with RILPL2 during ciliogenesis blockade

Herschel S Dhekne1, Izumi Yanatori1, Edmundo G Vides1

  • 1Department of Biochemistry, Stanford University School of Medicine, Stanford, CA, USA.

Life Science Alliance
|March 17, 2021
PubMed

Insights

Pathogenic LRRK2 kinase mutations in Parkinson's disease disrupt ciliogenesis by altering Rab GTPase phosphorylation. This study identifies RILPL2 and Myosin Va as key interactors that mislocalize, impairing cilia formation.

Area of Science:

  • Molecular biology
  • Cell biology
  • Neuroscience

Background:

  • Activating mutations in Leucine-rich repeat kinase 2 (LRRK2) are a significant genetic cause of Parkinson's disease.
  • Pathogenic LRRK2 phosphorylates specific Rab GTPases, leading to the disruption of ciliogenesis (the formation of cellular cilia).

Purpose of the Study:

  • To identify novel binding partners of LRRK2-phosphorylated Rab GTPases to elucidate the molecular mechanisms underlying LRRK2 pathogenesis.
  • To investigate the role of RILPL2 and Myosin Va in the context of LRRK2-mediated effects on ciliogenesis.

Main Methods:

  • Biochemical assays to determine binding affinities between RILPL2, Myosin Va, and phosphorylated Rab GTPases (Rab8A, Rab10).
  • Cell-based imaging techniques, including fluorescence loss in photobleaching microscopy, to track protein localization.
  • Analysis of ciliogenesis in cells with and without pathogenic LRRK2 activity and/or RILPL2 manipulation.

Main Results:

  • RILPL2 exhibits a strong preference for binding LRRK2-phosphorylated Rab8A and Rab10.
  • Myosin Va, a Rab effector, also binds LRRK2-phosphorylated Rab10 with high affinity.
  • Pathogenic LRRK2 causes phosphoRab10-dependent relocalization of RILPL2 and Myosin Va to the pericentriolar region, interfering with ciliogenesis.

Conclusions:

  • LRRK2-generated phosphoRab10 recruits RILPL2 and Myosin Va to the mother centriole, disrupting Myosin Va's function in ciliogenesis.
  • These findings highlight a novel mechanism by which LRRK2 mutations impair cilia formation, contributing to Parkinson's disease pathogenesis.

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