The In Situ Structure of Parkinson’s Disease-Linked LRRK2

• Published in: Cell 2020-09, Vol.182 (6), p.1508-1518.e16
• Main Authors: Watanabe, Reika, Buschauer, Robert, Böhning, Jan, Audagnotto, Martina, Lasker, Keren, Lu, Tsan-Wen, Boassa, Daniela, Taylor, Susan, Villa, Elizabeth
• Format: Article
• Language: English
• Subjects: correlative light and electron microscopy; cryo-electron tomography; Cryoelectron Microscopy - methods; Cytoplasm - metabolism; Electron Microscope Tomography - methods; GTP Phosphohydrolases - chemistry; GTP Phosphohydrolases - metabolism; HEK293 Cells; Humans; integrative modeling; kinase; leucine-rich repeat kinase; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 - chemistry; Microscopy, Electron, Transmission; microtubule; Microtubules - chemistry; Microtubules - metabolism; Models, Chemical; Mutation; Parkinson Disease - genetics; Parkinson Disease - metabolism; Parkinson Disease - pathology; Parkinson's disease; Phosphotransferases - chemistry; Phosphotransferases - metabolism; Protein Domains; subtomogram analysis; WD40 Repeats
• ISSN: 0092-8674; 1097-4172
• DOI: 10.1016/j.cell.2020.08.004

Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most frequent cause of familial Parkinson’s disease. LRRK2 is a multi-domain protein containing a kinase and GTPase. Using correlative light and electron microscopy, in situ cryo-electron tomography, and subtomogram analysis, we reveal a 14-Å structure of LRRK2 bearing a pathogenic mutation that oligomerizes as a right-handed double helix around microtubules, which are left-handed. Using integrative modeling, we determine the architecture of LRRK2, showing that the GTPase and kinase are in close proximity, with the GTPase closer to the microtubule surface, whereas the kinase is exposed to the cytoplasm. We identify two oligomerization interfaces mediated by non-catalytic domains. Mutation of one of these abolishes LRRK2 microtubule-association. Our work demonstrates the power of cryo-electron tomography to generate models of previously unsolved structures in their cellular environment. [Display omitted] •Cryo-electron tomography and integrative modeling reveal LRRK2’s structure in situ•Parkinson’s disease-linked LRRK2 forms double-helical filaments around microtubules•Homotypic interactions between the WD40 and COR domains form the filaments•The GTPase and kinase domains face the microtubule and cytoplasm, respectively Cryo-electron tomography combined with integrative modeling yields the structure of LRKK2 in a cellular context and shows how filaments of the protein embrace microtubules.