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FRET-based Tau seeding assay does not represent prion-like templated assembly of Tau filaments.

MPS-Authors
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Irsen,  Stephan
Electron Microscopy and Analytics, Center of Advanced European Studies and Research (caesar), Max Planck Society;

Klatt,  Christoph
Max Planck Research Group Structural Dynamics of Proteins, Center of Advanced European Studies and Research (caesar), Max Planck Society;

/persons/resource/persons128150

Mandelkow,  Eva-Maria
Neuronal Cytoskeleton and Alzheimer's Disease, Cooperations, Center of Advanced European Studies and Research (caesar), Max Planck Society;

/persons/resource/persons128150

Mandelkow,  Eva-Maria
Neuronal Cytoskeleton and Alzheimer's Disease, Cooperations, Center of Advanced European Studies and Research (caesar), Max Planck Society;

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s13024-020-00389-1.pdf
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Citation

Kaniyappan, S., Tepper, K., Biernat, J., Chandupatla, R. R., Hübschmann, S., Irsen, S., et al. (2020). FRET-based Tau seeding assay does not represent prion-like templated assembly of Tau filaments. Molecular neurodegeneration, 15(1): 39. doi:10.1186/s13024-020-00389-1.


Cite as: http://hdl.handle.net/21.11116/0000-0006-D0B8-F
Abstract
Tau aggregation into amyloid fibers based on the cross-beta structure is a hallmark of several Tauopathies, including Alzheimer Disease (AD). Trans-cellular propagation of Tau with pathological conformation has been suggested as a key disease mechanism. This is thought to cause the spreading of Tau pathology in AD by templated conversion of naive Tau in recipient cells into a pathological state, followed by assembly of pathological Tau fibers, similar to the mechanism of nucleated polymerization proposed for prion pathogenesis. In cell cultures, the process is often monitored by a FRET assay where the recipient cell expresses the Tau repeat domain (TauRD) with a pro-aggregant mutation, fused to GFP-based FRET pairs. Since the size of the reporter GFP (barrel of ~ 3 nm × 4 nm) is ~ 7 times larger than the β-strand distance (0.47 nm), this points to a potential steric clash. Hence, we investigated the influence of the GFP tag on TauFL or TauRD aggregation. Using biophysical methods (light scattering, atomic force microscopy (AFM), and scanning-transmission electron microscopy (STEM)), we found that the assembly of TauRD-GFP was severely inhibited and incompatible with that of Alzheimer filaments. These observations argue against the hypothesis that the propagation of Tau pathology in AD is caused by the prion-like templated aggregation of Tau protein, transmitted via cell-to-cell spreading of Tau. Thus, even though the observed local increase of FRET in recipient cells may be a valid hallmark of a pathological reaction, our data argue that it is caused by a process distinct from assembly of TauRD filaments.