Journal: Elife / Year: 2022 Title: Architecture of the chikungunya virus replication organelle. Authors: Timothée Laurent / Pravin Kumar / Susanne Liese / Farnaz Zare / Mattias Jonasson / Andreas Carlson / Lars-Anders Carlson / Abstract: are mosquito-borne viruses that cause serious disease in humans and other mammals. Along with its mosquito vector, the s chikungunya virus (CHIKV) has spread explosively in the last 20 years, and ... are mosquito-borne viruses that cause serious disease in humans and other mammals. Along with its mosquito vector, the s 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.
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