title: Novel ethyl methanesulfonate (EMS)-induced null alleles of the Drosophila homolog of LRRK2 reveal a crucial role in endolysosomal functions and autophagy in vivo.
abstract: Mutations in LRRK2 cause a dominantly inherited form of Parkinson's disease (PD) and are the most common known genetic determinant of PD. Inhibitor-based therapies targeting LRRK2 have emerged as a key therapeutic strategy in PD; thus, understanding the consequences of inhibiting the normal cellular functions of this protein is vital. Despite much interest, the physiological functions of LRRK2 remain unclear. Several recent studies have linked the toxicity caused by overexpression of pathogenic mutant forms of LRRK2 to defects in the endolysosomal and autophagy pathways, raising the question of whether endogenous LRRK2 might play a role in these processes. Here, we report the characterization of multiple novel ethyl methanesulfonate (EMS)-induced nonsense alleles in the Drosophila LRRK2 homolog, lrrk. Using these alleles, we show that lrrk loss-of-function causes striking defects in the endolysosomal and autophagy pathways, including the accumulation of markedly enlarged lysosomes that are laden with undigested contents, consistent with a defect in lysosomal degradation. lrrk loss-of-function also results in the accumulation of autophagosomes, as well as the presence of enlarged early endosomes laden with mono-ubiquitylated cargo proteins, suggesting an additional defect in lysosomal substrate delivery. Interestingly, the lysosomal abnormalities in these lrrk mutants can be suppressed by a constitutively active form of the small GTPase rab9, which promotes retromer-dependent recycling from late endosomes to the Golgi. Collectively, our data provides compelling evidence of a vital role for lrrk in lysosomal function and endolysosomal membrane transport in vivo, and suggests a link between lrrk and retromer-mediated endosomal recycling.
authors.affiliation: Department of Neurology, University of California, Los Angeles, CA 90095, USA. Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA.
authors.forename: Mark W
authors.initials: MW
authors.lastname: Dodson
authors.affiliation: Department of Neurology, University of California, Los Angeles, CA 90095, USA.
authors.forename: Lok K
authors.initials: LK
authors.lastname: Leung
authors.affiliation: Department of Neurology, University of California, Los Angeles, CA 90095, USA.
authors.forename: Mohiddin
authors.initials: M
authors.lastname: Lone
authors.affiliation: Department of Neurology, University of California, Los Angeles, CA 90095, USA. Brain Research Institute, The David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Lizzio
authors.affiliation: Department of Neurology, University of California, Los Angeles, CA 90095, USA. Molecular Biology Institute, University of California, Los Angeles, CA 90095, USA. Brain Research Institute, The David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA. Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095, USA. mingfly@ucla.edu.
authors.forename: Ming
authors.initials: M
authors.lastname: Guo
mesh_terms: D000483:Alleles; D000818:Animals; D030801:Animals, Genetically Modified; D001343:Autophagy; D018389:Codon, Nonsense; D003600:Cytosol / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics* / Q000502:physiology; D004331:Drosophila melanogaster; D011992:Endosomes / Q000378:metabolism; D005020:Ethyl Methanesulfonate / Q000737:chemistry*; D005260:Female; D006801:Humans; D020287:In Situ Nick-End Labeling; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D008247:Lysosomes / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009154:Mutation; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000502:physiology
chemical_list: D018389:Codon, Nonsense; D029721:Drosophila Proteins; D005020:Ethyl Methanesulfonate; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases; C495280:LRRK2 protein, human; C543697:LRRK protein, Drosophila
keywords: Autophagy; Drosophila; Endosomes; LRRK2; Lysosome; Parkinson’s disease; Rab7; Rab9