2026-06-14T06:44:30Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/69452025-04-01T09:57:30Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609
Repisalud
author
Rodriguez Martinez, Javier M
author
Chichón, Francisco Javier
author
Martin-Forero, Esther
author
Gonzalez-Camacho, Fernando
author
Carrascosa, José L
author
Castón, José R
author
Luque, Daniel
funder
Instituto de Salud Carlos III
2018-12-26T14:45:20Z
2018-12-26T14:45:20Z
2014-05-29
PLoS Pathog. 2014 May 29;10(5):e1004157
10.1371/journal.ppat.1004157
1553-7374
1553-7374
PLoS pathogens
24873828
http://hdl.handle.net/20.500.12105/6945
The infectivity of rotavirus, the main causative agent of childhood diarrhea, is dependent on activation of the extracellular viral particles by trypsin-like proteases in the host intestinal lumen. This step entails proteolytic cleavage of the VP4 spike protein into its mature products, VP8* and VP5*. Previous cryo-electron microscopy (cryo-EM) analysis of trypsin-activated particles showed well-resolved spikes, although no density was identified for the spikes in uncleaved particles; these data suggested that trypsin activation triggers important conformational changes that give rise to the rigid, entry-competent spike. The nature of these structural changes is not well understood, due to lack of data relative to the uncleaved spike structure. Here we used cryo-EM and cryo-electron tomography (cryo-ET) to characterize the structure of the uncleaved virion in two model rotavirus strains. Cryo-EM three-dimensional reconstruction of uncleaved virions showed spikes with a structure compatible with the atomic model of the cleaved spike, and indistinguishable from that of digested particles. Cryo-ET and subvolume average, combined with classification methods, resolved the presence of non-icosahedral structures, providing a model for the complete structure of the uncleaved spike. Despite the similar rigid structure observed for uncleaved and cleaved particles, trypsin activation is necessary for successful infection. These observations suggest that the spike precursor protein must be proteolytically processed, not to achieve a rigid conformation, but to allow the conformational changes that drive virus entry.
eng
New insights into rotavirus entry machinery: stabilization of rotavirus spike conformation is independent of trypsin cleavage
research article
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URL
https://repisalud.isciii.es/bitstreams/ee2a438a-740a-4f74-9a45-07f8c1b5f464/download
File
MD5
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application/pdf
NewInsightsIntoRotavirus_2014.pdf
URL
https://repisalud.isciii.es/bitstreams/72d7c619-ad48-4f13-840c-3737b6b6bdb9/download
File
MD5
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text/plain
NewInsightsIntoRotavirus_2014.pdf.txt