title: Insight into the mode of action of the LRRK2 Y1699C pathogenic mutant.
abstract: Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most prevalent known cause of autosomal dominant Parkinson's disease. The LRRK2 gene encodes a Roco protein featuring a Ras of complex proteins (ROC) GTPase and a kinase domain linked by the C-terminal of ROC (COR) domain. Here, we explored the effects of the Y1699C pathogenic LRRK2 mutation in the COR domain on GTPase activity and interactions within the catalytic core of LRRK2. We observed a decrease in GTPase activity for LRRK2 Y1699C comparable to the decrease observed for the R1441C pathogenic mutant and the T1348N dysfunctional mutant. To study the underlying mechanism, we explored the dimerization in the catalytic core of LRRK2. ROC-COR dimerization was significantly weakened by the Y1699C or R1441C/G mutation. Using a competition assay, we demonstrated that the intra-molecular ROC : COR interaction is favoured over ROC : ROC dimerization. Interestingly, the intra-molecular ROC : COR interaction was strengthened by the Y1699C mutation. This is supported by a 3D homology model of the ROC-COR tandem of LRRK2, showing that Y1699 is positioned at the intra-molecular ROC : COR interface. In conclusion, our data provides mechanistic insight into the mode of action of the Y1699C LRRK2 mutant: the Y1699C substitution, situated at the intra-molecular ROC : COR interface, strengthens the intra-molecular ROC : COR interaction, thereby locally weakening the dimerization of LRRK2 at the ROC-COR tandem domain resulting in decreased GTPase activity.
authors.affiliation: Laboratory for Neurobiology and Gene Therapy, Division of Molecular Medicine, Department of Molecular and Cellular Medicine, Katholieke Universiteit Leuven, Leuven, Belgium.
authors.forename: Veronique
authors.initials: V
authors.lastname: Daniëls
authors.forename: Renée
authors.initials: R
authors.lastname: Vancraenenbroeck
authors.forename: Bernard M H
authors.initials: BM
authors.lastname: Law
authors.forename: Elisa
authors.initials: E
authors.lastname: Greggio
authors.forename: Evy
authors.initials: E
authors.lastname: Lobbestael
authors.forename: Fangye
authors.initials: F
authors.lastname: Gao
authors.forename: Marc
authors.initials: M
authors.lastname: De Maeyer
authors.forename: Mark R
authors.initials: MR
authors.lastname: Cookson
authors.forename: Kirsten
authors.initials: K
authors.lastname: Harvey
authors.forename: Veerle
authors.initials: V
authors.lastname: Baekelandt
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Taymans
mesh_terms: D020134:Catalytic Domain / Q000235:genetics; D003545:Cysteine / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D009154:Mutation; D011485:Protein Binding / Q000235:genetics; D055503:Protein Multimerization / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D014443:Tyrosine / Q000235:genetics
chemical_list: D014443:Tyrosine; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases; D020558:GTP Phosphohydrolases; D003545:Cysteine