April 2020 Nsp3 abcam ab181620 Nucleocapsid Sino Biological 40143-MM05 Validation Commercial antibodies validated as per manufacturers website: Beta actin Sigma A5441 Immunoblot on chicken fibroblast cell extracts Spike Abcam Ab252690 Validated by ELISA on free peptide from SARS-CoV-1 Nsp3 abcam ab181620 Validated by western blot on SARS-CoV-1 infected cells Nucleocapsid Sino Biological 40143-MM05 Validated by western blot with corresponding viruses Eukaryotic cell lines Policy information about cell lines Cell line source(s) VeroE6 cells were obtained from ATCC, Calu-3 cells were obtained from Manfred Frey, originally from ATCC. Authentication Cells were not further authenticated Mycoplasma contamination Cells have been tested and are free of mycoplasma.

No abstract

The ordered assembly of tau protein into abnormal filamentous inclusions underlies many human neurodegenerative diseases. Tau assemblies seem to spread through specific neural networks in each disease, with short filaments having the greatest seeding activity. The abundance of tau inclusions strongly correlates with disease symptoms. Six tau isoforms are expressed in the normal adult human brain-three isoforms with four microtubule-binding repeats each (4R tau) and three isoforms that lack the second repeat (3R tau). In various diseases, tau filaments can be composed of either 3R or 4R tau, or of both. Tau filaments have distinct cellular and neuroanatomical distributions, with morphological and biochemical differences suggesting that they may be able to adopt disease-specific molecular conformations. Such conformers may give rise to different neuropathological phenotypes, reminiscent of prion strains. However, the underlying structures are not known. Using electron cryo-microscopy, we recently reported the structures of tau filaments from patients with Alzheimer's disease, which contain both 3R and 4R tau. Here we determine the structures of tau filaments from patients with Pick's disease, a neurodegenerative disorder characterized by frontotemporal dementia. The filaments consist of residues Lys254-Phe378 of 3R tau, which are folded differently from the tau filaments in Alzheimer's disease, establishing the existence of conformers of assembled tau. The observed tau fold in the filaments of patients with Pick's disease explains the selective incorporation of 3R tau in Pick bodies, and the differences in phosphorylation relative to the tau filaments of Alzheimer's disease. Our findings show how tau can adopt distinct folds in the human brain in different diseases, an essential step for understanding the formation and propagation of molecular conformers.

Methods are presented that detect three types of aberrations in single-particle cryo-EM data sets: symmetrical and antisymmetrical optical aberrations and magnification anisotropy. Because these methods only depend on the availability of a preliminary 3D reconstruction from the data, they can be used to correct for these aberrations for any given cryo-EM data set, a posteriori. Using five publicly available data sets, it is shown that considering these aberrations improves the resolution of the 3D reconstruction when these effects are present. The methods are implemented in version 3.1 of the open-source software package RELION. research papers IUCrJ (2020). 7, 253-267 Jasenko Zivanov et al. High-order aberrations and anisotropic magnification 255