Structural Correlates of Rotavirus Cell Entry

Aliaa H. Abdelhakim, Eric N. Salgado, Xiaofeng Fu, Mithun Pasham, Daniela Nicastro, Tomas Kirchhausen, Stephen C. Harrison

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Cell entry by non-enveloped viruses requires translocation into the cytosol of a macromolecular complex—for double-strand RNA viruses, a complete subviral particle. We have used live-cell fluorescence imaging to follow rotavirus entry and penetration into the cytosol of its ∼700 Å inner capsid particle (“double-layered particle”, DLP). We label with distinct fluorescent tags the DLP and each of the two outer-layer proteins and track the fates of each species as the particles bind and enter BSC-1 cells. Virions attach to their glycolipid receptors in the host cell membrane and rapidly become inaccessible to externally added agents; most particles that release their DLP into the cytosol have done so by ∼10 minutes, as detected by rapid diffusional motion of the DLP away from residual outer-layer proteins. Electron microscopy shows images of particles at various stages of engulfment into tightly fitting membrane invaginations, consistent with the interpretation that rotavirus particles drive their own uptake. Electron cryotomography of membrane-bound virions also shows closely wrapped membrane. Combined with high resolution structural information about the viral components, these observations suggest a molecular model for membrane disruption and DLP penetration.

Original languageEnglish (US)
JournalPLoS pathogens
Volume10
Issue number9
DOIs
StatePublished - Sep 1 2014

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ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Molecular Biology
  • Genetics
  • Virology

Cite this

Abdelhakim, A. H., Salgado, E. N., Fu, X., Pasham, M., Nicastro, D., Kirchhausen, T., & Harrison, S. C. (2014). Structural Correlates of Rotavirus Cell Entry. PLoS pathogens, 10(9). https://doi.org/10.1371/journal.ppat.1004355