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- Title
A Bayesian approach to single-particle electron cryo-tomography in RELION-4.0.
- Authors
Zivanov, Jasenko; Otón, Joaquín; Zunlong Ke; von Kügelgen, Andriko; Pyle, Euan; Kun Qu; Morado, Dustin; Castaño-Díez, Daniel; Zanetti, Giulia; Bharat, Tanmay A. M.; Briggs, John A. G.; Scheres, Sjors H. W.
- Abstract
We present a new approach for macromolecular structure determination from multiple particles in electron cryo-tomography (cryo-ET) data sets. Whereas existing subtomogram averaging approaches are based on 3D data models, we propose to optimise a regularised likelihood target that approximates a function of the 2D experimental images. In addition, analogous to Bayesian polishing and contrast transfer function (CTF) refinement in single-particle analysis, we describe the approaches that exploit the increased signal-to-noise ratio in the averaged structure to optimise tilt-series alignments, beam-induced motions of the particles throughout the tilt-series acquisition, defoci of the individual particles, as well as higher-order optical aberrations of the microscope. Implementation of our approaches in the open-source software package RELION aims to facilitate their general use, particularly for those researchers who are already familiar with its single-particle analysis tools. We illustrate for three applications that our approaches allow structure determination from cryo-ET data to resolutions sufficient for de novo atomic modelling.
- Subjects
OPTICAL aberrations; ATOMIC models; ELECTRONS; PARTICLE motion; OPTICAL microscopes
- Publication
eLife, 2023, p1
- ISSN
2050-084X
- Publication type
Article
- DOI
10.7554/eLife.83724