The<i>in situ</i>structure of Parkinson’s disease-linked LRRK2

Watanabe, Reika; Buschauer, Robert; Böhning, Jan; Audagnotto, Martina; Lasker, Keren; Lu, Tsan Wen; Boassa, Daniela; Taylor, Susan S.; Villa, Elizabeth

doi:10.1101/837203

Cited by 25 publications

(61 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, a 14Å structure of microtubule-associated filaments of fulllength LRRK2 (carrying the filament-promoting I2020T mutation 13 ) was reported using in situ cryo-ET and sub-tomogram averaging 21 (Fig. 2a).…”

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

“…2a). The LRRK2 filaments formed on microtubules were right-handed 21 . The fact that microtubules are left-handed, combined with the absence of Fig.…”

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

“…h, i, The helix has dimensions compatible with the diameter of a 12-protofilament microtubule (EMD-5192) 53 , which was the species used to obtain the tomographic reconstruction shown in Fig. 2a 21 , and has its RoC domains pointing towards the microtubule surface. strong density connecting the LRRK2 filament to the microtubule surface 21 raised the question of whether LRRK2's interaction with microtubules is direct or is mediated by other microtubule-associated proteins.…”

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

“…2a 21 , and has its RoC domains pointing towards the microtubule surface. strong density connecting the LRRK2 filament to the microtubule surface 21 raised the question of whether LRRK2's interaction with microtubules is direct or is mediated by other microtubule-associated proteins. To address this, we combined purified microtubules with purified LRRK2 RCKW and observed them by cryo-EM.…”

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

“…Previously, integrative modeling was used to build a model into the in situ structure of microtubule-associated LRRK2 21 . This modeling indicated that the observed density was comprised of the RoC, COR, Kinase and WD40 domains and gave orientation ensembles for each domain 21 that were in good agreement with the high-resolution structure of LRRK2 RCKW presented here.…”

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

See 4 more Smart Citations

Parkinson’s Disease-linked LRRK2 structure and model for microtubule interaction

Salogiannis

Mathea

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Leucine Rich Repeat Kinase 2 (LRRK2) is the most commonly mutated gene in familial Parkinson's disease. LRRK2 is proposed to function in membrane trafficking and co-localizes with microtubules. We report the 3.5Å structure of the catalytic half of LRRK2, and an atomic model of microtubule-associated LRRK2 built using a reported 14Å cryo-electron tomography in situ structure. We propose that the conformation of LRRK2's kinase domain regulates its microtubule interaction, with a closed conformation favoring binding. We show that the catalytic half of LRRK2 is sufficient for microtubule binding and blocks the motility of the microtubule-based motors kinesin and dynein in vitro. Kinase inhibitors that stabilize an open conformation relieve this interference and reduce LRRK2 filament formation in cells, while those that stabilize a closed conformation do not. Our findings suggest that LRRK2 is a roadblock for microtubule-based motors and have implications for the design of therapeutic LRRK2 kinase inhibitors.

show abstract

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

“…2a). The LRRK2 filaments formed on microtubules were right-handed 21 . The fact that microtubules are left-handed, combined with the absence of Fig.…”

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

Section: An Atomic Model Of Microtubule-bound Lrrk2 Filamentsmentioning

confidence: 99%

See 3 more Smart Citations

Parkinson’s Disease-linked LRRK2 structure and model for microtubule interaction

Salogiannis

Mathea

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

The promise and the challenges of cryo‐electron tomography

2020

View full text Add to dashboard Cite

Structural biologists have traditionally approached cellular complexity in a reductionist manner in which the cellular molecular components are fractionated and purified before being studied individually. This ‘divide and conquer’ approach has been highly successful. However, awareness has grown in recent years that biological functions can rarely be attributed to individual macromolecules. Most cellular functions arise from their concerted action, and there is thus a need for methods enabling structural studies performed in situ, ideally in unperturbed cellular environments. Cryo‐electron tomography (Cryo‐ET) combines the power of 3D molecular‐level imaging with the best structural preservation that is physically possible to achieve. Thus, it has a unique potential to reveal the supramolecular architecture or ‘molecular sociology’ of cells and to discover the unexpected. Here, we review state‐of‐the‐art Cryo‐ET workflows, provide examples of biological applications, and discuss what is needed to realize the full potential of Cryo‐ET.

show abstract

The Dynamo Software Package for Cryo-electron Tomography and Subtomogram Averaging

et al. 2020

View full text Add to dashboard Cite

Cryo-electron tomography (cryo-ET) and subtomogram averaging (STA) have become popular techniques for structural determination of biological macrocomplexes in their native environment. Cryo-ET allows three-dimensional (3D) visualization of cellular organelles and macromolecular complexes at nanometer resolution 1 in their close-to-native environment. Briefly, in cryo-ET, the specimen is physically rotated around a single axis inside the electron microscope. This axis is perpendicular to the optical axis of the microscope, and thus, to the direction of the electron beam. Micrographs are collected at each rotation angle, resulting in a set of tilted micrographs referred to as tilt series. Each micrograph represents a view of the specimen at different tilt angles. The tilt series is then computationally aligned and reconstructed into a 3D tomogram. Unfortunately, the rotation possible, i.e., the tilt range, is physically restricted by the microscope geometry, thus, tilt series are usually acquired from-60° to +60°. As a result, reconstructed tomograms experience partial sampling or missing information in Fourier Space. The missing information exhibits a characteristic wedge-like shape, which is known as the missing wedge. In real space, the missing wedge manifests itself as a blurred elongation of the imaged specimen in the direction of the electron beam 2 .

show abstract

Thein situstructure of Parkinson’s disease-linked LRRK2

Cited by 25 publications

References 59 publications

Parkinson’s Disease-linked LRRK2 structure and model for microtubule interaction

Parkinson’s Disease-linked LRRK2 structure and model for microtubule interaction

The promise and the challenges of cryo‐electron tomography

The Dynamo Software Package for Cryo-electron Tomography and Subtomogram Averaging

Contact Info

Product

Resources

About