Abstract
Mutations in the gene coding for the multi-domain protein leucine-rich repeat kinase 2 (LRRK2) are the leading cause of genetically inherited Parkinson’s disease (PD). Two of the common found mutations are the R1441C and G2019S. In this study we identified protein phosphatase 2A (PP2A) as an interacting partner of LRRK2. We were able to demonstrate that the Ras of complex protein (ROC) domain is sufficient to interact with the three subunits of PP2A in human neuroblastoma SH-SY5Y cells and in HeLa cells. The alpha subunit of PP2A is interacting with LRRK2 in the perinuclear region of HeLa cells. Silencing the catalytic subunit of PP2A by shRNA aggravated cellular degeneration induced by the pathogenic R1441C-LRRK2 mutant expressed in neuroblastoma SH-SY5Y cells. A similar enhancement of apoptotic nuclei was observed by downregulation of the catalytic subunit of PP2A in cultured cortical cells derived from neurons overexpressing the pathogenic mutant G2019S-LRRK2. Conversely, pharmacological activation of PP2A by sodium selenate showed a partial neuroprotection from R1441C-LRRK2-induced cellular degeneration. All these data suggest that PP2A is a new interacting partner of LRRK2 and reveal the importance of PP2A as a potential therapeutic target in PD.
Acknowledgments
The research leading to these results has received funding from the European Community’s Framework Programme (FP7/2007–2013) under grant agreement number TRANSPOL – 264399 and by FP6 ITN INTCHEM. We would like to thank Sabine Laerbusch, Dini Nurul Binte Abdul Rahim and Sally Dong Qianying for the technical help, and Dr. Nadine Ackermann, Dr. Daniela Damen, Dr. Koushik Chakrabarty, Dr. Anja Ehrkamp and Dr. Veena Nambiar Potheraveedu for the inspiring discussions. We would also like to thank Prof. Dr. Bernhard Hovemann and Dr. Christoph Goemanns for providing us with the LRRK2 and H2B-RFP plasmids, respectively. We would also like to thank Prof. Steve Finkbeiner (Gladstone Institutes, US), for providing us with the pGW1-Venus and pGW1-Venus WT LRRK2 plasmids. Furthermore we would like to thank Dr. Arjan Kortholt (University of Groningen, The Netherlands) for providing us with the GFP-ROC-COR plasmid (human sequence). Finally we would like to thank Dr. Benjamin Franzel for his help in establishing the mass spectrometric data and Mrs Semra Ince and Mr. Sergii Shydlovskyi for their help and suggestions regarding the GTP binding assay. We thank RUBION for providing facilities. Finally, we would like to acknowledge Dr. Jian Zhong and Dr. Dianna Willis for their helpful suggestions. The authors declare that they have no conflict of interest. All persons gave their informed consent prior to their inclusion in the study. All animal studies have been approved by the appropriate Ethics Committee and have therefore been performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments.
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Article note:
This article was submitted for the Highlight Issue ‘Perspectives of Molecular Neuroscience in Health and Disease’, published in issue 3 (2016) of Biological Chemistry.
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The online version of this article (DOI: 10.1515/hsz-2015-0189) offers supplementary material, available to authorized users.
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