Cryo-Electron Tomography: The Resolution Revolution and a Surge of In Situ Virological Discoveries

Abstract

The recent proliferation of cryo-electron tomography (cryo-ET) techniques has led to the cryo-ET resolution revolution. Meanwhile, significant efforts have been made to improve the identification of targets in the cellular context and the throughput of cryo-focused ion beam (FIB) milling. Together, these developments led to a surge of in situ discoveries on how enveloped viruses are assembled and how viruses interact with cells in infected host. In this article, we review the recent advances in cryo-ET, high-resolution insights into virus assembly, and the findings from inside infected eukaryotic and prokaryotic cells. Expected final online publication date for the Annual Review of Biophysics, Volume 52 is May 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Recent advances in infectious disease research using cryo-electron tomography

“…Cryo-ET is rapidly maturing as a technology, catalyzing significant advancements in our understanding of viral infection biology. Several groups have recently reviewed cryo-ET advances in studying viral infections ( Graham and Zhang, 2023 ; Hong et al, 2023 ; Zimmermann and Chlanda, 2023 ) .…”

MemBrain v2: an end-to-end tool for the analysis of membranes in cryo-electron tomography

“…Cryo-ET visualizes all densities inside a vitreously frozen cellular volume, creating vast potential to investigate the structures, interactions, and spatial arrangements of diverse organellar and macromolecular components. However, this biological complexity is also a central challenge for analyzing cryo-ET data (Young and Villa, 2023), and it remains difficult to accurately annotate the many features inside cellular tomograms. Membranes and their embedded protein complexes are fundamental to numerous biological processes that are visible by cryo-ET, including the ER-Golgi secretory pathway (Gemmer et al, 2023; Albert et al, 2020; Tran et al, 2021; Bykov et al, 2017), autophagy (M. Li et al, 2023; Bieber et al, 2022), vesicular transport (Foster et al, 2021), synaptic transmission (Radecke et al, 2023; Tao et al, 2018; Radhakrishnan et al, 2021; Liu, Tao, et al, 2020), energy generation in mitochondrial cristae and chloroplast thylakoids (Barad et al, 2023; Waltz et al, 2021; Huokko et al, 2021; Wietrzynski et al, 2020; Gupta et al, 2021), organelle contact sites (Wozny et al, 2023; Collado et al, 2019), cell membrane protrusions (Rodrigues-Oliveira et al, 2023; Sartori-Rupp et al, 2019), viral replication (Mendonça et al, 2021; Klein, Cortese, et al, 2020; Wolff et al, 2020; Klein, Golani,et al, 2023; Armbruster et al, 2023; Dahmane et al, 2022), and bacterial cell division (Navarro et al, 2022; Khanna et al, 2021).…”

Rapid structural analysis of bacterial ribosomesin situ

“…In the last decade, cryogenic focused ion beam milling (cryo-FIB) in conjunction with cryo-electron tomography (cryo-ET) has revolutionized structural biology [1][2][3][4][5][6][7] . Indeed, when combined with subtomogram averaging (STA), cryo-ET can produce structures at resolutions supporting molecular interpretations (~3-10 Å), and it allows one to chart the dynamic landscapes of macromolecular complexes [8][9][10][11] .…”