Mutations in the LRRK2 Roc-COR tandem domain link Parkinson's disease to Wnt signaling pathways

Department of Pharmacology, The School of Pharmacy, Brunswick Square, London, UK.
Human Molecular Genetics (Impact Factor: 6.39). 08/2009; 18(20):3955-68. DOI: 10.1093/hmg/ddp337
Source: PubMed


Mutations in PARK8, encoding LRRK2, are the most common known cause of Parkinson's disease. The LRRK2 Roc-COR tandem domain exhibits GTPase activity controlling LRRK2 kinase activity via an intramolecular process. We report the interaction of LRRK2 with the dishevelled family of phosphoproteins (DVL1-3), key regulators of Wnt (Wingless/Int) signalling pathways important for axon guidance, synapse formation and neuronal maintenance. Interestingly, DVLs can interact with and mediate the activation of small GTPases with structural similarity to the LRRK2 Roc domain. The LRRK2 Roc-COR domain and the DVL1 DEP domain were necessary and sufficient for LRRK2-DVL1 interaction. Co-expression of DVL1 increased LRRK2 steady-state protein levels, an effect that was dependent on the DEP domain. Strikingly, LRRK2-DVL1-3 interactions were disrupted by the familial PARK8 mutation Y1699C, whereas pathogenic mutations at residues R1441 and R1728 strengthened LRRK2-DVL1 interactions. Co-expression of DVL1 with LRRK2 in mammalian cells resulted in the redistribution of LRRK2 to typical cytoplasmic DVL1 aggregates in HEK293 and SH-SY5Y cells and co-localization in neurites and growth cones of differentiated dopaminergic SH-SY5Y cells. This is the first report of the modulation of a key LRRK2-accessory protein interaction by PARK8 Roc-COR domain mutations segregating with Parkinson's disease. Since the DVL1 DEP domain is known to be involved in the regulation of small GTPases, we propose that: (i) DVLs may influence LRRK2 GTPase activity, and (ii) Roc-COR domain mutations modulating LRRK2-DVL interactions indirectly influence kinase activity. Our findings also link LRRK2 to Wnt signalling pathways, suggesting novel pathogenic mechanisms and new targets for genetic analysis in Parkinson's disease.

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    • "The studies showing LRRK2 interacting with many varied cellular proteins present a challenge for understanding its function. LRRK2 has been suggested to bind to many different proteins, including moesin, tubulin, MKK3, 6 and 7, JIP1, 3, and 4, arfGAP1, arhGEF7, endoA, cyclin-GAK, rab5, rab7L1, 14-3-3, (Imai et al., 2008; Shin et al., 2008; Ko et al., 2009; Sancho et al., 2009; Gehrke et al., 2010; Hsu et al., 2010a,b; Kumar et al., 2010; Nichols et al., 2010; Chan et al., 2011; Matta et al., 2012; Stafa et al., 2012; Habig et al., 2013; Beilina et al., 2014). LRRK2 has a regulatory role in a wide variety of biological processes; such as, protein translations, cytoskeletal processes, vesicular dynamics, neurite extension, mitochondrial function, endoplasmic reticulum function, and autophagy. "
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    ABSTRACT: LRRK2 is a protein that interacts with a plethora of signaling molecules, but the complexity of LRRK2 function presents a challenge for understanding the role of LRRK2 in the pathophysiology of Parkinson's disease (PD). Studies of LRRK2 using over-expression in transgenic mice have been disappointing, however, studies using invertebrate systems have yielded a much clearer picture, with clear effects of LRRK2 expression, knockdown or deletion in Caenorhabditis elegans and Drosophila on modulation of survival of dopaminergic neurons. Recent studies have begun to focus attention on particular signaling cascades that are a target of LRRK2 function. LRRK2 interacts with members of the mitogen activated protein kinase (MAPK) pathway and might regulate the pathway action by acting as a scaffold that directs the location of MAPK pathway activity, without strongly affecting the amount of MAPK pathway activity. Binding to GTPases, GTPase-activating proteins and GTPase exchange factors are another strong theme in LRRK2 biology, with LRRK2 binding to rac1, cdc42, rab5, rab7L1, endoA, RGS2, ArfGAP1, and ArhGEF7. All of these molecules appear to feed into a function output for LRRK2 that modulates cytoskeletal outgrowth and vesicular dynamics, including autophagy. These functions likely impact modulation of α-synuclein aggregation and associated toxicity eliciting the disease processes that we term PD.
    Full-text · Article · Jul 2014 · Frontiers in Molecular Neuroscience
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    • "In addition, LRRK2 has been reported to interact with various small GTPases including Rac1 (Chan et al., 2011), rab7L1/rab29 (MacLeod et al., 2013), rab5b (Shin et al., 2008), rab7 (Dodson et al., 2012), and with a GEF and a GAP, respectively (Haebig et al., 2010; Stafa et al., 2012; Xiong et al., 2012). Other interactors, such as components of a membrane-associated Wnt signalling complex (Sancho et al., 2009; Berwick and Harvey, 2012a), clathrin (Daechsel et al., 2007; Piccoli et al., 2011), clathrin adaptor proteins (AP-1, AP-2) and accessory proteins (dynamin and AP180) (Piccoli et al., 2011; Stafa et al., 2013), or the LRRK2-mediated regulation of membrane association of endophilinA via phosphorylation (Matta et al., 2012) all indicate that LRRK2 may play an important role in multiple vesicular trafficking steps such as synaptic vesicle recycling, receptor internalization, early and late endocytic trafficking, endosome- Golgi trafficking and autophagy (see below). "
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    ABSTRACT: Mutations in LRRK2 (leucine-rich repeat kinase 2) are found associated with both sporadic and familial Parkinson´s disease (PD). Pathogenic mutations are localized to the catalytic domains of LRRK2, including kinase and GTPase domains. Altered catalytic activity correlates with neurotoxicity, indicating that targeting those activities may provide clues as to novel therapeutic strategies for LRRK2-linked PD. However, the cellular readout of such altered catalytic activities remains largely unknown. Recent cell biological studies have started to highlight possible early cellular events which are altered in the presence of pathogenic LRRK2 and may ultimately lead to neuronal demise, and these studies link altered LRRK2 function to various abnormal endolysosomal vesicular trafficking events. This review examines our current knowledge of LRRK2 neurobiology and how pathogenic mutations may lead to neurodegeneration in PD.
    Full-text · Article · May 2014 · Neuropharmacology
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    • "LRRK2 has been linked to a multitude of cellular functions and pathways, including regulation of neurite outgrowth, Wnt signaling, mitochondrial disease, and autophagy (Dächsel et al., 2010; Winner et al., 2011; Berwick and Harvey, 2012; Papkovskaia et al., 2012). Several studies have identified interaction partners of LRRK2, including 14-3-3, Tubulin, ArfGAP1, Rac1, and DVL (Sancho et al., 2009; Chan et al., 2011; Kawakami et al., 2012; Xiong et al., 2012; Dzamko et al., 2013; Fraser et al., 2013). Despite all this accumulating data, substantial gaps remain in the knowledge about the underlying pathways of LRRK2 mediated PD. "
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    ABSTRACT: Human leucine rich repeat kinase 2 (LRRK2) belongs to the Roco family of proteins, which are characterized by the presence of a Ras-like G-domain (Roc), a C-terminal of Roc domain (COR), and a kinase domain. Mutations in LRRK2 have been found to be thus far the most frequent cause of late-onset Parkinson's disease (PD). Several of the pathogenic mutations in LRRK2 result in decreased GTPase activity and enhanced kinase activity, suggesting a possible PD-related gain of abnormal function. Important progress in the structural understanding of LRRK2 has come from our work with related Roco proteins from lower organisms. Atomic structures of Roco proteins from prokaryotes revealed that Roco proteins belong to the GAD class of molecular switches (G proteins activated by nucleotide dependent dimerization). As in LRRK2, PD-analogous mutations in Roco proteins from bacteria decrease the GTPase reaction. Studies with Roco proteins from the model organism Dictyostelium discoideum revealed that PD mutants have different effects and most importantly they explained the G2019S-related increased LRRK2 kinase activity. Furthermore, the structure of Dictyostelium Roco4 kinase in complex with the LRRK2 inhibitor H1152 showed that Roco4 and other Roco family proteins can be important for the optimization of the current, and identification of new, LRRK2 kinase inhibitors. In this review we highlight the recent progress in structural and biochemical characterization of Roco proteins and discuss its implication for the understanding of the complex regulatory mechanism of LRRK2.
    Full-text · Article · May 2014 · Frontiers in Molecular Neuroscience
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