The WD40 Domain Is Required for LRRK2 Neurotoxicity

Abstract

BackgroundMutations in leucine-rich repeat kinase 2 (LRRK2) are the most common genetic cause of Parkinson disease (PD). LRRK2 contains an “enzymatic core” composed of GTPase and kinase domains that is flanked by leucine-rich repeat (LRR) and WD40 protein-protein interaction domains. While kinase activity and GTP-binding have both been implicated in LRRK2 neurotoxicity, the potential role of other LRRK2 domains has not been as extensively explored.Principal FindingsWe demonstrate that LRRK2 normally exists in a dimeric complex, and that removing the WD40 domain prevents complex formation and autophosphorylation. Moreover, loss of the WD40 domain completely blocks the neurotoxicity of multiple LRRK2 PD mutations.ConclusionThese findings suggest that LRRK2 dimerization and autophosphorylation may be required for the neurotoxicity of LRRK2 PD mutations and highlight a potential role for the WD40 domain in the mechanism of LRRK2-mediated cell death.

Heterogeneity of Leucine-Rich Repeat Kinase 2 Mutations: Genetics, Mechanisms and Therapeutic Implications

“…The substitution of a neutral and flexible glycine for a positively charged arginine at this position could interfere with interdomain interactions within LRRK2. The WD40 and extreme C-terminus of LRRK2 are also required for kinase activity of LRRK2 as C-terminally truncated constructs have a loss of kinase activity [67,73,103]. In our hands, using a range of different constructs and assays, the G2385R substitution causes a 40-50 % loss of LRRK2 kinase function.…”

“…Overexpression of G2019S LRRK2 in cultured primary neurons and neuroblastoma cell lines mediates cellular toxicity and triggers apoptotic cell death [13,15,70,103]. Similarly, overexpression of R1441C or Y1699C also triggers cell death [13].…”

“…I2020T also induces the shortening and debranching of neurites [56,126]. Acute overexpression of R1441C LRRK2 induces neurite shortening [13,56,70,103]. Primary neurons derived from bacterial artificial chromosome transgenic (Y1699C) mice have reduced neurite outgrowth and branching, which was partially rescued with staurosporine, a nonspecific kinase inhibitor [136].…”

LRRK2 Controls Synaptic Vesicle Storage and Mobilization within the Recycling Pool

“…Data obtained from our domain-based interaction studies finally suggest that LRRK2 interacts with presynaptic partners mainly through its WD40 C-terminal domain. This domain is required for both LRRK2 toxic and physiological role (Jorgensen et al, 2009;Sheng et al, 2010), and it harbors the mutation G2385R, considered as the main risk factor for Parkinson's disease in Chinese Han population . Future studies are now needed to determine whether perturbed regulation of vesicle trafficking may contribute to Parkinson's disease associated with this gene variant.…”

Receptor-interacting protein (RIP) kinase family

“…73 The kinase activity, GTP-binding (GTPase) and WD40 domain of RIP7 are all reported to be implicated in RIP7's neurotoxicity. [74][75][76][77][78] Although much effort has been made in searching for the molecular mechanism underlying RIP7's function, [79][80][81][82][83][84][85] the pathogenesis of RIP7 mutations in Parkinson's disease remains unclear.…”