Phosphatome-wide siRNA screen reveals phosphorylation regulators of the Parkinson’s disease protein LRRK2.

Abstract

Event Abstract Back to Event Phosphatome-wide siRNA screen reveals phosphorylation regulators of the Parkinson’s disease protein LRRK2. Tina De Wit1*, Evy Lobbestael1, Marc Böllliger2, Matthieu Drouyer3, Marie-Christine Chartier-Harlin3, R. Jeremy Nichols2, Jean-Marc Taymans3 and Veerle Baekelandt1 1 KU Leuven, Belgium 2 The Parkinson's Institute, United States 3 UMR-S1172 - Jean-Pierre Aubert Research Center, France Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common known cause of inherited Parkinson’s disease. Interestingly, cellular phosphorylation levels on S910/S935/S955/S973 of LRRK2 appear to be strongly regulated and might be important for the LRRK2 pathomechanism, as these are dephosphorylated in several pathogenic mutants (R1441G, Y1699C and I2020T). We have shown previously that protein phosphatase 1 (PP1) recruitment and PP1-mediated dephosphorylation underlie the reduced LRRK2 phosphorylation observed in the R1441G mutant and strikingly, also upon LRRK2 kinase inhibitor treatment, a potential therapy for PD. From an siRNA screen against 298 phosphatase targets in U2OS cells expressing LRRK2-eGFP, we selected 39 proteins with the greatest effect on LRRK2 S935 phosphorylation level. These were further validated using lentiviral vector-mediated miRNA-based knock-down or overexpression in SH-SY5Y cells and HEK293T cells stably overexpressing LRRK2. Preliminary data point to a role for both PP1 and PP2A, together with some of their regulatory proteins, as important phosphatases for the LRRK2 phosphorylation cycle. Currently, we are further validating a selection of PP1 and PP2A holoenzymes for their role in LRRK2 phosphorylation regulation in basal conditions and after LRRK2 kinase inhibition. Identification of the catalytic and/or regulatory subunits of phosphatases that are involved in LRRK2 dephosphorylation, will give insight in kinase inhibitor-mediated effects and may lead to new therapies for PD. Keywords: Parkinson’s disease, LRRK2, Phosphatases, siRNA, LRRK2 kinase inhibition Conference: 12th National Congress of the Belgian Society for Neuroscience, Gent, Belgium, 22 May - 22 May, 2017. Presentation Type: Poster Presentation Topic: Disorders of the Nervous System Citation: De Wit T, Lobbestael E, Böllliger M, Drouyer M, Chartier-Harlin M, Nichols R, Taymans J and Baekelandt V (2019). Phosphatome-wide siRNA screen reveals phosphorylation regulators of the Parkinson’s disease protein LRRK2.. Front. Neurosci. Conference Abstract: 12th National Congress of the Belgian Society for Neuroscience. doi: 10.3389/conf.fnins.2017.94.00094 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 21 Apr 2017; Published Online: 25 Jan 2019. * Correspondence: Miss. Tina De Wit, KU Leuven, Leuven, Belgium, tina.dewit@kuleuven.be Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tina De Wit Evy Lobbestael Marc Böllliger Matthieu Drouyer Marie-Christine Chartier-Harlin R. Jeremy Nichols Jean-Marc Taymans Veerle Baekelandt Google Tina De Wit Evy Lobbestael Marc Böllliger Matthieu Drouyer Marie-Christine Chartier-Harlin R. Jeremy Nichols Jean-Marc Taymans Veerle Baekelandt Google Scholar Tina De Wit Evy Lobbestael Marc Böllliger Matthieu Drouyer Marie-Christine Chartier-Harlin R. Jeremy Nichols Jean-Marc Taymans Veerle Baekelandt PubMed Tina De Wit Evy Lobbestael Marc Böllliger Matthieu Drouyer Marie-Christine Chartier-Harlin R. Jeremy Nichols Jean-Marc Taymans Veerle Baekelandt Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.