Abstract
Understanding how viruses with multi-segmented genomes incorporate one copy of each segment into their capsids remains an intriguing question. Here, we review our recent progress and describe the advancements made in understanding the genome packaging mechanism of a model nonenveloped virus, Bluetongue virus (BTV), with a 10-segment (S1–S10) double-strand RNA (dsRNA) genome. BTV (multiple serotypes), a member of the Orbivirus genus in the Reoviridae family, is a notable pathogen for livestock and is responsible for significant economic losses worldwide. This has enabled the creation of an extensive set of reagents and assays, including reverse genetics, cell-free RNA packaging, and bespoke bioinformatics approaches, which can be directed to address the packaging question. Our studies have shown that (i) UTRs enable the conformation of each segment necessary for the next level of RNA–RNA interaction; (ii) a specific order of intersegment interactions leads to a complex RNA network containing all the active components in sorting and packaging; (iii) networked segments are recruited into nascent assembling capsids; and (iv) select capsid proteins might be involved in the packaging process. The key features of genome packaging mechanisms for BTV and related dsRNA viruses are novel and open up new avenues of potential intervention.
Highlights
Genome packaging is an essential process in the virus life cycle, as at least one genome must be incorporated into the limited space available in the capsid
We have summarized how multipartite genomic ssRNA segments are combined prior to, or during, packaging using an experimental model virus, Bluetongue virus (BTV), with direct implications for related medically important viruses (e.g., Rotaviruses) and many other complex double-stranded RNA (dsRNA)
An extensive background knowledge coupled with multidisciplinary approaches, including reverse genetics and a cell-free
Summary
Genome packaging is an essential process in the virus life cycle, as at least one genome (or one set of RNAs of a multi-segmented genome) must be incorporated into the limited space available in the capsid. 2).,these thesepackaged packaged ssRNAs serve leading theformation formationof ofaa stable stable core ssRNAs serve as templates for dsRNA synthesis, resulting in an equimolar set of all genome segments. These in vitro assembled cores, with a complete set of genomic RNA segments, are replication competent, but how the 10 ssRNAs are recognised precisely and packaged. 3, x FOR PEER REVIEW as templates for dsRNA synthesis, resulting in an equimolar set of all genome segments These in vitro assembled cores, with a complete set of genomic RNA segments, are replication competent, but how the 10 ssRNAs are recognised precisely and packaged with the correct stoichiometry has been unclear until recently. BTV cores (indicated arrows)micrograph [21]
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