Abstract
Mutations in LRRK2 cause familial Parkinson’s disease and common variants increase disease risk. LRRK2 kinase activity and cellular localization are tightly regulated by phosphorylation of key residues, primarily Ser1292 and Ser935, which impacts downstream phosphorylation of its substrates, among which Rab10. A comprehensive characterization of LRRK2 activity and phosphorylation in brain as a function of age and mutations is missing. Here, we monitored Ser935 and Ser1292 phosphorylation in midbrain, striatum, and cortex of 1, 6, and 12 months-old mice carrying G2019S and R1441C mutations or murine bacterial artificial chromosome (BAC)-Lrrk2-G2019S. We observed that G2019S and, at a greater extent, R1441C brains display decreased phospho-Ser935, while Ser1292 autophosphorylation increased in G2019S but not in R1441C brain, lung, and kidney compared to wild-type. Further, Rab10 phosphorylation, is elevated in R1441C carrying mice, indicating that the effect of LRRK2 mutations on substrate phosphorylation is not generalizable. In BAC-Lrrk2-G2019S striatum and midbrain, Rab10 phosphorylation, but not Ser1292 autophosphorylation, decreases at 12-months, pointing to autophosphorylation and substrate phosphorylation as uncoupled events. Taken together, our study provides novel evidence that LRRK2 phosphorylation in mouse brain is differentially impacted by mutations, brain area, and age, with important implications as diagnostic markers of disease progression and stratification.
Highlights
Mutations in the gene encoding leucine-rich repeat kinase 2 (LRRK2) cause autosomal dominantParkinson’s disease (PD), while common variants in the LRRK2 locus increase the lifetime risk of disease [1]
LRRK2 phosphorylation in mouse brain, we dissected threeregions, brain regions, To explore LRRK2 phosphorylation in mouse brain, we dissected three brain namely namely striatum, cortex, and midbrain
Because lungs and kidneys have been previously shown to express high levels of LRRK2 [19,20,21] and LRRK2 inhibitors cause morphological changes in type II pneumocytes [22], we evaluated LRRK2 phosphorylation in these tissues isolated from 12 months-old WT, R1441C knock-in (RCKI), G2019S knock-in (GSKI), and Lrrk2 KO mice
Summary
Mutations in the gene encoding leucine-rich repeat kinase 2 (LRRK2) cause autosomal dominantParkinson’s disease (PD), while common variants in the LRRK2 locus increase the lifetime risk of disease [1]. Mutations in the gene encoding leucine-rich repeat kinase 2 (LRRK2) cause autosomal dominant. LRRK2 is a large signaling protein comprising serine-threonine kinase and ROC-COR. Cells 2020, 9, 2344 function through a direct activating effect on kinase activity (G2019S) or by decreasing GTP hydrolysis (R1441C/G/H and Y1699C), which affects LRRK2 subcellular access to its substrates, the most validated being a subset of Rab GTPases [3]. Within a cluster of N-terminus residues, the two major ones being Ser910 and Ser935 (reviewed in [4]). Phosphorylation of these residues provides the docking sites for the binding of 14-3-3 proteins, which regulate LRRK2 activity and subcellular localization [5,6]. LRRK2 kinase inhibition results in LRRK2 dephosphorylation at Ser910/Ser935 and consequent dissociation from 14-3-3 proteins [4,5,6]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
