3D Cryo-Correlative Methods to Study Virus Structure and Dynamics Within Cells.

Abstract

Understanding the dynamic processes involving virus structural components within host cells is crucial for comprehending viral infection, as viruses rely entirely on host cells for replication. Viral infection involves various intracellular stages, including cell entry, genome uncoating, replication, transcription and translation, assembly of new virus particles in a complex morphogenetic process, and the release of new virions from the host cell. These events are dynamic and scarce and can be obscured by other cellular processes, necessitating novel approaches for their in situ characterization. Among these methods, correlative microscopy integrates the labeling, localization, and functional characterization of events of interest through visible light microscopy, complemented by the structural insights provided by high-resolution imaging techniques. This correlative approach enables a comprehensive exploration of subcellular events within the cellular context, including those related to viral morphogenesis. This chapter provides an introduction to correlative three-dimensional imaging methods, specifically designed to study viral morphogenesis and other intracellular stages of the viral cycle under conditions closely resembling their native environment. The integration of whole-cell imaging and high-resolution structural biology techniques is emphasized as essential for unraveling the mechanisms by which viruses generate and disseminate their progeny.