title: Protection against Mitochondrial and Metal Toxicity Depends on Functional Lipid Binding Sites in ATP13A2.
abstract: The late endo-/lysosomal P-type ATPase ATP13A2 (PARK9) is implicated in Parkinson's disease (PD) and Kufor-Rakeb syndrome, early-onset atypical Parkinsonism. ATP13A2 interacts at the N-terminus with the signaling lipids phosphatidic acid (PA) and phosphatidylinositol (3,5) bisphosphate (PI(3,5)P2), which modulate ATP13A2 activity under cellular stress conditions. Here, we analyzed stable human SHSY5Y cell lines overexpressing wild-type (WT) or ATP13A2 mutants in which three N-terminal lipid binding sites (LBS1-3) were mutated. We explored the regulatory role of LBS1-3 in the cellular protection by ATP13A2 against mitochondrial stress induced by rotenone and found that the LBS2-3 mutants displayed an abrogated protective effect. Moreover, in contrast to WT, the LBS2 and LBS3 mutants responded poorly to pharmacological inhibition of, respectively, PI(3,5)P2 and PA formation. We further demonstrate that PA and PI(3,5)P2 are also required for the ATP13A2-mediated protection against the toxic metals Mn(2+), Zn(2+), and Fe(3+), suggesting a general lipid-dependent activation mechanism of ATP13A2 in various PD-related stress conditions. Our results indicate that the ATP13A2-mediated protection requires binding of PI(3,5)P2 to LBS2 and PA to LBS3. Thus, targeting the N-terminal lipid binding sites of ATP13A2 might offer a therapeutic approach to reduce cellular toxicity of various PD insults including mitochondrial stress.
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium; Laboratory of Cell Death Research and Therapy, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium.
authors.forename: Shaun
authors.initials: S
authors.lastname: Martin
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium.
authors.forename: Sarah
authors.initials: S
authors.lastname: van Veen
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium.
authors.forename: Tine
authors.initials: T
authors.lastname: Holemans
authors.affiliation: Laboratory of Cell Death Research and Therapy, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium.
authors.forename: Seyma
authors.initials: S
authors.lastname: Demirsoy
authors.affiliation: Research Group for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Kapucijnenvoer 33, Blok i, P.O. Box 7001, 3000 Leuven, Belgium; Leuven Viral Vector Core, KU Leuven, Kapucijnenvoer 33, Blok i, P.O. Box 7001, 3000 Leuven, Belgium.
authors.forename: Chris
authors.initials: C
authors.lastname: van den Haute
authors.affiliation: Research Group for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven, Kapucijnenvoer 33, Blok i, P.O. Box 7001, 3000 Leuven, Belgium.
authors.forename: Veerle
authors.initials: V
authors.lastname: Baekelandt
authors.affiliation: Laboratory of Cell Death Research and Therapy, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Agostinis
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Eggermont
authors.affiliation: Laboratory of Cellular Transport Systems, Department of Cellular and Molecular Medicine, KU Leuven, Campus Gasthuisberg O&N 1, Herestraat 49, P.O. Box 802, 3000 Leuven, Belgium.
authors.forename: Peter
authors.initials: P
authors.lastname: Vangheluwe