A Bayesian approach to single-particle electron cryo-tomography in RELION-4.0

Zivanov, J. and Otón, J. and Ke, Z. and von Kügelgen, A. and Pyle, E. and Qu, K. and Morado, D. and Castano-Daez, D. and Zanetti, Giulia and Bharat, T.A.M. and Briggs, J.A.G. and Scheres, S.H.W. (2023) A Bayesian approach to single-particle electron cryo-tomography in RELION-4.0. eLife 11 , ISSN 2050-084X.

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.

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