Using cryoEM and cryoET to visualize membrane penetration of a non-enveloped virus

Xian Xia; Z Hong Zhou

doi:10.1016/j.xpro.2022.101825

Using cryoEM and cryoET to visualize membrane penetration of a non-enveloped virus

STAR Protoc. 2022 Dec 16;3(4):101825. doi: 10.1016/j.xpro.2022.101825. Epub 2022 Nov 11.

Authors

Xian Xia¹, Z Hong Zhou²

Affiliations

¹ Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
² Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, 90095, USA; California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA. Electronic address: hong.zhou@ucla.edu.

Abstract

Key to cell entry by non-enveloped viruses is virus-cell interactions at the cell or endosomal membrane. Here, we detail our protocols to capture such interactions between non-enveloped virus bluetongue virus (BTV) and vesicular membrane by cryogenic electron microscopy (cryoEM) and tomography (cryoET). Key steps include virus isolation, liposome preparation, virus-liposome incubation and vitrification, cryoEM and cryoET imaging, data processing for 3D reconstruction, and subtomogram averaging. The protocols can be generally applicable to studies of cell entry by other non-enveloped viruses. For complete details on the use and execution of this protocol, please refer to Xia et al. (2021).

Keywords: Cryo-EM; Microbiology; Microscopy; Structural biology.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural

MeSH terms

Animals
Cryoelectron Microscopy / methods
Endosomes
Liposomes*
Microscopy, Electron
Viruses*

Substances

Liposomes

Abstract

Publication types

MeSH terms

Substances

Grants and funding