Rotavirus genomic RNA complex forms via specific RNA–RNA interactions: Disruption of RNA complex inhibits virus infectivity

Teodoro Fajardo, Po Yu Sung, Cristina Celma, Polly Roy

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

Rotavirus (RV), a member of the Reoviridae family, causes infection in children and infants, with high morbidity and mortality. To be viable, the virus particle must package a set of eleven RNA segments. In order to understand the packaging mechanism, here, we co-synthesized sets of RNA segments in vitro in different combinations and detected by two alternate methods: the electrophoretic mobility shift assay (EMSA) and the RNA-bead pull-down assay. We showed that viral positive-sense RNA segments interact with each other in a specific manner, forming RNA complexes, and that the RNA–RNA interactions followed a sequential order initiated by small RV segments. Further, we demonstrated that RNA complexes were perturbed by targeted specific antisense oligoribonucleotides (ORNs) complementary to short RNA sequences, indicating that the RNA–RNA interactions between different segments were sequence-specific. The same inhibitory ORNs also had the capability to inhibit virus replication. The combined in vitro and in vivo data inferred that RNA–RNA interactions and specific complex formation are essential for sorting different segments, possibly prior to, or during, genome packaging. As genome assembly is a universal requirement in the Reoviridae family, this work offers an approach towards a further understanding of the sorting and packaging mechanisms of RV and related dsRNA (double-stranded RNA) viruses.

Original languageEnglish
Article number167
JournalViruses
Volume9
Issue number7
DOIs
Publication statusPublished - Jul 2017
Externally publishedYes

Keywords

  • Genome packaging
  • Reoviridae
  • RNA interaction
  • Rotavirus
  • Virus assembly

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