14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity
LRRK2 mutations are among the most common genetic causes for Parkinson’s disease (PD), and toxicity is associated with increased kinase activity. 14-3-3 proteins are key interactors that regulate LRRK2 kinase activity. Phosphorylation of the 14-3-3θ isoform at S232 is dramatically increased in human...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Cold Spring Harbor Laboratory
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10312468/ https://www.ncbi.nlm.nih.gov/pubmed/37398189 http://dx.doi.org/10.1101/2023.05.27.542591 |
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author | Pattanayak, Rudradip Petit, Chad M. Yacoubian, Talene A. |
author_facet | Pattanayak, Rudradip Petit, Chad M. Yacoubian, Talene A. |
author_sort | Pattanayak, Rudradip |
collection | PubMed |
description | LRRK2 mutations are among the most common genetic causes for Parkinson’s disease (PD), and toxicity is associated with increased kinase activity. 14-3-3 proteins are key interactors that regulate LRRK2 kinase activity. Phosphorylation of the 14-3-3θ isoform at S232 is dramatically increased in human PD brains. Here we investigate the impact of 14-3-3θ phosphorylation on its ability to regulate LRRK2 kinase activity. Both wildtype and the non-phosphorylatable S232A 14-3-3θ mutant reduced the kinase activity of wildtype and G2019S LRRK2, whereas the phosphomimetic S232D 14-3-3θ mutant had minimal effects on LRRK2 kinase activity, as determined by measuring autophosphorylation at S1292 and T1503 and Rab10 phosphorylation. However, wildtype and both 14-3-3θ mutants similarly reduced the kinase activity of the R1441G LRRK2 mutant. 14-3-3θ phosphorylation did not promote global dissociation with LRRK2, as determined by co-immunoprecipitation and proximal ligation assays. 14-3-3s interact with LRRK2 at several phosphorylated serine/threonine sites, including T2524 in the C-terminal helix, which can fold back to regulate the kinase domain. Interaction between 14-3-3θ and phosphorylated T2524 LRRK2 was important for 14-3-3θ’s ability to regulate kinase activity, as wildtype and S232A 14-3-3θ failed to reduce the kinase activity of G2019S/T2524A LRRK2. Molecular modeling showed that 14-3-3θ phosphorylation causes a partial rearrangement of its canonical binding pocket, thus affecting the interaction between 14-3-3θ and the C-terminus of LRRK2. We conclude that 14-3-3θ phosphorylation destabilizes the interaction of 14-3-3θ with LRRK2 at T2524, which consequently promotes LRRK2 kinase activity. |
format | Online Article Text |
id | pubmed-10312468 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-103124682023-07-01 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity Pattanayak, Rudradip Petit, Chad M. Yacoubian, Talene A. bioRxiv Article LRRK2 mutations are among the most common genetic causes for Parkinson’s disease (PD), and toxicity is associated with increased kinase activity. 14-3-3 proteins are key interactors that regulate LRRK2 kinase activity. Phosphorylation of the 14-3-3θ isoform at S232 is dramatically increased in human PD brains. Here we investigate the impact of 14-3-3θ phosphorylation on its ability to regulate LRRK2 kinase activity. Both wildtype and the non-phosphorylatable S232A 14-3-3θ mutant reduced the kinase activity of wildtype and G2019S LRRK2, whereas the phosphomimetic S232D 14-3-3θ mutant had minimal effects on LRRK2 kinase activity, as determined by measuring autophosphorylation at S1292 and T1503 and Rab10 phosphorylation. However, wildtype and both 14-3-3θ mutants similarly reduced the kinase activity of the R1441G LRRK2 mutant. 14-3-3θ phosphorylation did not promote global dissociation with LRRK2, as determined by co-immunoprecipitation and proximal ligation assays. 14-3-3s interact with LRRK2 at several phosphorylated serine/threonine sites, including T2524 in the C-terminal helix, which can fold back to regulate the kinase domain. Interaction between 14-3-3θ and phosphorylated T2524 LRRK2 was important for 14-3-3θ’s ability to regulate kinase activity, as wildtype and S232A 14-3-3θ failed to reduce the kinase activity of G2019S/T2524A LRRK2. Molecular modeling showed that 14-3-3θ phosphorylation causes a partial rearrangement of its canonical binding pocket, thus affecting the interaction between 14-3-3θ and the C-terminus of LRRK2. We conclude that 14-3-3θ phosphorylation destabilizes the interaction of 14-3-3θ with LRRK2 at T2524, which consequently promotes LRRK2 kinase activity. Cold Spring Harbor Laboratory 2023-05-30 /pmc/articles/PMC10312468/ /pubmed/37398189 http://dx.doi.org/10.1101/2023.05.27.542591 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Pattanayak, Rudradip Petit, Chad M. Yacoubian, Talene A. 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity |
title | 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity |
title_full | 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity |
title_fullStr | 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity |
title_full_unstemmed | 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity |
title_short | 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate LRRK2 kinase activity |
title_sort | 14-3-3 phosphorylation inhibits 14-3-3θ’s ability to regulate lrrk2 kinase activity |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10312468/ https://www.ncbi.nlm.nih.gov/pubmed/37398189 http://dx.doi.org/10.1101/2023.05.27.542591 |
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