Abstract
Parkinson's disease predisposing LRRK2 kinase phosphorylates a group of Rab GTPase proteins including Rab29, within the effector‐binding switch II motif. Previous work indicated that Rab29, located within the PARK16 locus mutated in Parkinson's patients, operates in a common pathway with LRRK2. Here, we show that Rab29 recruits LRRK2 to the trans‐Golgi network and greatly stimulates its kinase activity. Pathogenic LRRK2 R1441G/C and Y1699C mutants that promote GTP binding are more readily recruited to the Golgi and activated by Rab29 than wild‐type LRRK2. We identify conserved residues within the LRRK2 ankyrin domain that are required for Rab29‐mediated Golgi recruitment and kinase activation. Consistent with these findings, knockout of Rab29 in A549 cells reduces endogenous LRRK2‐mediated phosphorylation of Rab10. We show that mutations that prevent LRRK2 from interacting with either Rab29 or GTP strikingly inhibit phosphorylation of a cluster of highly studied biomarker phosphorylation sites (Ser910, Ser935, Ser955 and Ser973). Our data reveal that Rab29 is a master regulator of LRRK2, controlling its activation, localization, and potentially biomarker phosphorylation.
Highlights
Autosomal dominant missense mutations within the leucine-rich repeat protein kinase 2 (LRRK2) gene predispose to Parkinson’s disease (Paisan-Ruiz et al, 2004; Zimprich et al, 2004)
There are 14 Rab proteins that are phosphorylated by LRRK2 (Steger et al, 2017) and conceivably, LRRK2 mutants may have slightly different localization or preferences for diverse Rab proteins, which could account for variation between Ser1292 phosphorylation and Rab10 phosphorylation observed (Fig 1A)
We found that stimulation of Ser1292 as well as Rab10 phosphorylation induced by overexpression of Rab29 was abolished by introducing a kinase-inactivating D2017A mutation (Fig 1A, right panels), confirming that Rab29 was enhancing phosphorylation by stimulating LRRK2 kinase activity
Summary
Autosomal dominant missense mutations within the leucine-rich repeat protein kinase 2 (LRRK2) gene predispose to Parkinson’s disease (Paisan-Ruiz et al, 2004; Zimprich et al, 2004). Pathogenic mutations located within the GTPase (R1441G/C) and COR (Y1699C) domains do not directly stimulate LRRK2 kinase activity in vitro (Jaleel et al, 2007; Nichols et al, 2010); they markedly enhance phosphorylation of Rab isoforms to an even greater extent than the G2019S mutation in vivo (Ito et al, 2016; Steger et al, 2016). These mutations promote GTP binding to the LRRK2 Roc domain (Guo et al, 2007; Lewis et al, 2007; Li et al, 2007; Daniels et al, 2011; Webber et al, 2011; Liao et al, 2014).
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