Lysosomal positioning regulates Rab10 phosphorylation at LRRK2 ⁺ lysosomes

Abstract

Genetic variation at the leucine-rich repeat kinase 2 ( LRRK2 ) locus contributes to an enhanced risk of familial and sporadic Parkinson’s disease. Previous data have demonstrated that recruitment to various membranes of the endolysosomal system results in LRRK2 activation. However, the mechanism(s) underlying LRRK2 activation at endolysosomal membranes and the cellular consequences of these events are still poorly understood. Here, we directed LRRK2 to lysosomes and early endosomes, triggering both LRRK2 autophosphorylation and phosphorylation of the direct LRRK2 substrates Rab10 and Rab12. However, when directed to the lysosomal membrane, pRab10 was restricted to perinuclear lysosomes, whereas pRab12 was visualized on both peripheral and perinuclear LRRK2 + lysosomes, suggesting that lysosomal positioning provides additional regulation of LRRK2-dependent Rab phosphorylation. Anterograde transport of lysosomes to the cell periphery by increasing the expression of ARL8B and SKIP or by knockdown of JIP4 blocked the recruitment and phosphorylation of Rab10 by LRRK2. The absence of pRab10 from the lysosomal membrane prevented the formation of a lysosomal tubulation and sorting process we previously named LYTL. Conversely, overexpression of RILP resulted in lysosomal clustering within the perinuclear area and increased LRRK2-dependent Rab10 recruitment and phosphorylation. The regulation of Rab10 phosphorylation in the perinuclear area depends on counteracting phosphatases, as the knockdown of phosphatase PPM1H significantly increased pRab10 signal and lysosomal tubulation in the perinuclear region. Our findings suggest that LRRK2 can be activated at multiple cellular membranes, including lysosomes, and that lysosomal positioning further provides the regulation of some Rab substrates likely via differential phosphatase activity or effector protein presence in nearby cellular compartments.

Axonal transport of autophagosomes is regulated by dynein activators JIP3/JIP4 and ARF/RAB GTPases

“…Notably, both JIP1 and JIP3/4 have been previously implicated in the transport of RAB10+ vesicles (Kluss et al, 2022; Bonet-Ponce et al, 2020; Deng et al, 2014). In our study, RAB10 appears to increase the recruitment and/or activation of kinesin on LC3+ and LAMP1+ puncta.…”

“…In our study, RAB10 appears to increase the recruitment and/or activation of kinesin on LC3+ and LAMP1+ puncta. RAB10, like many other RABs, is regulated by phosphorylation with phospho-RAB10 (especially T73) being generally more active and membrane-associated (Yan et al, 2018; Lara Ordóñez et al, 2022; Kluss et al, 2022; Wauters et al, 2020; Waschbüsch et al, 2020; Homma et al, 2021). RAB10, especially phospho-RAB10, regulates the motility of multiple kinesin-1 and kinesin-3 cargoes within cells (Etoh and Fukuda, 2019; Deng et al, 2014; Taylor et al, 2015; Zajac and Horne-Badovinac, 2022).…”

“…Interestingly, overexpression of GTP-locked ARF6 is sufficient to ameliorate the AV motility phenotype observed in hyperactive LRRK2 mutant conditions (Dou et al, 2022). Under increased LRRK2 activity, JIP3/4 seem to exhibit enhanced interaction with kinesin and phosphorylated RABs (Dou et al, 2022; Boecker et al, 2021; Kluss et al, 2022; Bonet-Ponce et al, 2020). In these conditions, GTP-locked ARF6 expression presumably scaffolds the formation of more JIP3/4-dynein-dynactin complexes to compete with the RAB-JIP3/4-kinesin-1 complexes (Dou et al, 2022).…”

microRNAs profiling of small extracellular vesicles from midbrain tissue of Parkinson’s disease

“…However, neuron-astrocyte coupling of fatty acid metabolism is essential for neuron survival (Ioannou et al, 2019;Qi et al, 2021), and fatty acid metabolism of neurons is also of great importance. As for the predicted target genes of these miRNAs, the target gene SCYL3 is involved in maintaining motor neuron viability (Kuliyev et al, 2018); eIF4G2 is involved in neuropathic pain (Zhang et al, 2021); and Rab10, NF-1, and Dll1 are indeed involved in PD (Imai et al, 2015;Kuo et al, 2021;Kluss et al, 2022). Many studies have demonstrated that these four miRNAs are involved in regulating brain function or brain damage in humans, mice, and rats.…”

Dissecting the effects of GTPase and kinase domain mutations on LRRK2 endosomal localization and activity

“…Though largely cytoplasmic, LRRK2 can associate with vesicular membranes to phosphorylate membrane-bound Rabs 14–17 . In some conditions, such as prolonged lysosomal damage, LRRK2 has been shown to be retained on intracellular membranes to a level that can be readily observed by microscopy 16,18–21 . How different LRRK2 mutants affect its membrane association and Rab phosphorylation remains poorly characterized.…”