Characterization of the rotavirus assembly pathway in situ using cryoelectron tomography.

Autor:	Shah PNM; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, Headington, Oxford, UK; CAMS Oxford Institute, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Headington, Oxford, UK. Electronic address: pranav.shah@strubi.ox.ac.uk., Gilchrist JB; Diamond Light Source Ltd, Harwell Science & Innovation Campus, Didcot, UK., Forsberg BO; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, Headington, Oxford, UK; Department of Physiology and Pharmacology, Karolinska Institute, Stockholm, Sweden., Burt A; Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge, UK., Howe A; Diamond Light Source Ltd, Harwell Science & Innovation Campus, Didcot, UK., Mosalaganti S; Life Sciences Institute and Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, USA., Wan W; Vanderbilt University Center for Structural Biology, PMB 407917, 465 21st Ave S, 5140 MRB3, Nashville, TN, USA., Radecke J; Diamond Light Source Ltd, Harwell Science & Innovation Campus, Didcot, UK., Chaban Y; Diamond Light Source Ltd, Harwell Science & Innovation Campus, Didcot, UK., Sutton G; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, Headington, Oxford, UK., Stuart DI; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, Headington, Oxford, UK; CAMS Oxford Institute, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Headington, Oxford, UK; Diamond Light Source Ltd, Harwell Science & Innovation Campus, Didcot, UK. Electronic address: david.stuart@strubi.ox.ac.uk., Boyce M; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, Headington, Oxford, UK. Electronic address: mark.boyce@strubi.ox.ac.uk.
Jazyk:	angličtina
Zdroj:	Cell host & microbe [Cell Host Microbe] 2023 Apr 12; Vol. 31 (4), pp. 604-615.e4. Date of Electronic Publication: 2023 Mar 29.
DOI:	10.1016/j.chom.2023.03.004
Abstrakt:	Rotavirus assembly is a complex process that involves the stepwise acquisition of protein layers in distinct intracellular locations to form the fully assembled particle. Understanding and visualization of the assembly process has been hampered by the inaccessibility of unstable intermediates. We characterize the assembly pathway of group A rotaviruses observed in situ within cryo-preserved infected cells through the use of cryoelectron tomography of cellular lamellae. Our findings demonstrate that the viral polymerase VP1 recruits viral genomes during particle assembly, as revealed by infecting with a conditionally lethal mutant. Additionally, pharmacological inhibition to arrest the transiently enveloped stage uncovered a unique conformation of the VP4 spike. Subtomogram averaging provided atomic models of four intermediate states, including a pre-packaging single-layered intermediate, the double-layered particle, the transiently enveloped double-layered particle, and the fully assembled triple-layered virus particle. In summary, these complementary approaches enable us to elucidate the discrete steps involved in forming an intracellular rotavirus particle. Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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