Year: 2022
Type: article
Abstract: Cryo-electron microscopy has become an essential tool to understand structure and function of biological samples. Especially for pathogens, such as disease-causing bacteria and viruses, insights gained by cryo-EM can aid in developing cures. However, due to the biosafety restrictions of pathogens, samples are often treated by chemical fixation to render the pathogen inert, affecting the ultrastructure of the sample. Alternatively, researchers use in vitro or ex vivo models, which are non-pathogenic but lack the complexity of the pathogen of interest. Here we show that ultraviolet-C (UVC) radiation applied at cryogenic temperatures can be used to eliminate or dramatically reduce the infectivity of Vibrio cholerae and the bacterial virus, the ICP1 bacteriophage. We show no discernable structural impact of this treatment of either sample using two cryo-EM methods: cryo-electron tomography followed by sub-tomogram averaging, and single particle analysis (SPA). Additionally, we applied the UVC irradiation to the protein apoferritin (ApoF), which is a widely used test sample for high-resolution SPA studies. The UVC-treated ApoF sample resulted in a 2.1 Å structure indistinguishable from an untreated published map. This research demonstrates that UVC treatment is an effective and inexpensive addition to the cryo-EM sample preparation toolbox.
Source: Communications Biology
Language: English
Subfield: Structural Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,490
Funders: U.S. Department of Health and Human Services, Research Councils UK, European Commission, Nederlandse Organisatie voor Wetenschappelijk Onderzoek, National Institutes of Health, Directorate for Biological Sciences, Horizon 2020 Framework Programme, Biotechnology and Biological Sciences Research Council, National Institute of Allergy and Infectious Diseases, Center for Information Technology
Awards: G1464318188, G2374514745, Releasing the full potential of Instruct to expand and consolidate infrastructure services for integrated structural life science research, G2771723404, G4134265268, Assembly and Dynamics of Bacterial Chemosensory Signaling Arrays, G4699088171, G5255093015, Center for the Structural Biology of Cellular Host Elements in Egress, Trafficking, and Assembly of HIV (CHEETAH Center)
OpenAlex ID: W4220769548