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J Biol Chem. 2019 Mar 29;294(13):4738-4758. doi: 10.1074/jbc.RA118.005008. Epub 2019 Feb 1.

The G2019S variant of leucine-rich repeat kinase 2 (LRRK2) alters endolysosomal trafficking by impairing the function of the GTPase RAB8A.

Author information

1
From the Institute of Parasitology and Biomedicine "López-Neyra," Consejo Superior de Investigaciones Científicas (CSIC), Avda del Conocimiento s/n, 18016 Granada, Spain.
2
the Department of Pharmacology, Physiology and Neuroscience, New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, New Jersey 07103, and.
3
the Department of Biology, University of Padova, Padova 35121, Italy.
4
From the Institute of Parasitology and Biomedicine "López-Neyra," Consejo Superior de Investigaciones Científicas (CSIC), Avda del Conocimiento s/n, 18016 Granada, Spain, sabine.hilfiker@ipb.csic.es.

Abstract

Mutations in the gene encoding for leucine-rich repeat kinase 2 (LRRK2) are a common cause of hereditary Parkinson's disease. LRRK2 regulates various intracellular vesicular trafficking pathways, including endolysosomal degradative events such as epidermal growth factor receptor (EGFR) degradation. Recent studies have revealed that a subset of RAB proteins involved in secretory and endocytic recycling are LRRK2 kinase substrates in vivo However, the effects of LRRK2-mediated phosphorylation of these substrates on membrane trafficking remain unknown. Here, using an array of immunofluorescence and pulldown assays, we report that expression of active or phosphodeficient RAB8A variants rescues the G2019S LRRK2-mediated effects on endolysosomal membrane trafficking. Similarly, up-regulation of the RAB11-Rabin8-RAB8A cascade, which activates RAB8A, also reverted these trafficking deficits. Loss of RAB8A mimicked the effects of G2019S LRRK2 on endolysosomal trafficking and decreased RAB7A activity. Expression of pathogenic G2019S LRRK2 or loss of RAB8A interfered with EGFR degradation by causing its accumulation in a RAB4-positive endocytic compartment, which was accompanied by a deficit in EGFR recycling and was rescued upon expression of active RAB7A. Dominant-negative RAB7A expression resulted in similar deficits in EGF degradation, accumulation in a RAB4 compartment, and deficits in EGFR recycling, which were all rescued upon expression of active RAB8A. Taken together, these findings suggest that, by impairing RAB8A function, pathogenic G2019S LRRK2 deregulates endolysosomal transport and endocytic recycling events.

KEYWORDS:

GTPase; Parkinson disease; RAB7A; RAB8A; Rab; early recycling compartment; endolysosome; leucine-rich repeat kinase 2 (LRRK2); lysosome; neurodegeneration; protein homeostasis; receptor endocytosis; receptor recycling

PMID:
30709905
PMCID:
PMC6442034
DOI:
10.1074/jbc.RA118.005008
[Indexed for MEDLINE]
Free PMC Article

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