Architecture of the chikungunya virus replication organelle
- Umeå University, Sweden
- Max Planck Institute for the Physics of Complex Systems, Germany
- University of Oslo, Norway
Abstract
Alphaviruses are mosquito-borne viruses that cause serious disease in humans and other mammals. Along with its mosquito vector, the Alphavirus chikungunya virus (CHIKV) has spread explosively in the last 20 years, and there is no approved treatment for chikungunya fever. On the plasma membrane of the infected cell, CHIKV generates dedicated organelles for viral RNA replication, so-called spherules. Whereas structures exist for several viral proteins that make up the spherule, the architecture of the full organelle is unknown. Here, we use cryo-electron tomography to image CHIKV spherules in their cellular context. This reveals that the viral protein nsP1 serves as a base for the assembly of a larger protein complex at the neck of the membrane bud. Biochemical assays show that the viral helicase-protease nsP2, while having no membrane affinity on its own, is recruited to membranes by nsP1. The tomograms further reveal that full-sized spherules contain a single copy of the viral genome in double-stranded form. Finally, we present a mathematical model that explains the membrane remodeling of the spherule in terms of the pressure exerted on the membrane by the polymerizing RNA, which provides a good agreement with the experimental data. The energy released by RNA polymerization is found to be sufficient to remodel the membrane to the characteristic spherule shape.
Data availability
The subtomogram averages of the neck complex have been deposited at the Electron Microscopy Data Bank with accession codes EMD-14686 (unsymmetrized) and EMD-14687 (C12-symmetrized). Two reconstructed tomograms of CHIKV spherules at the plasma membrane, binned by a factor 4, are also available with the accession codes EMD-15582 and EMD-15583.
Article and author information
Author details
Lars-Anders Carlson
Funding
Human Frontier Science Program (CDA00047/2017-C)
- Lars-Anders Carlson
Vetenskapsrådet (2018-05851)
- Lars-Anders Carlson
Vetenskapsrådet (2021-01145)
- Lars-Anders Carlson
Kempestiftelserna (JCK-1723.2)
- Pravin Kumar
Max Planck Institute for the Physics of Complex Systems (open access funding)
- Susanne Liese
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2022, Laurent et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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