Name
The role of RNA-RNA interactions in selective genome packaging in rotavirus
Presenter
Aidan Tollervey, University of Cambridge
Co-Author(s)
Aidan Tollervey
Abstract Category
Virus Replication: Entry, Exit and Everything in Between
Abstract
The eleven-segmented rotavirus genome must undergo stoichiometric assortment of distinct RNA segments during replication. Multiplexed RNA FISH analysis suggests that all eleven RNAs accumulate in viroplasms, which are biomolecular condensates essential for rotaviral replication, yet the molecular basis of selective RNA packaging remains poorly understood. While sequence-specific protein–RNA interactions, such as VP1 polymerase binding to the 3′ UTR of rotaviral transcripts, are well characterised, accumulating evidence points to a critical role for RNA–RNA interactions in driving segment assortment.
To define these interactions, we applied RNA crosslinking approaches capable of capturing intermolecular RNA contacts, to enable us to dissect the impact of co-transcriptional folding and to compare RNA interaction networks formed by in vitro transcription versus those formed during in situ (intra-virion) transcription. In parallel, we are developing a live-cell imaging system using reverse genetics to rescue viruses suitable for real-time transcriptional imaging, offering temporal resolution beyond that of fixed-cell methods like single-molecule FISH.
Together, these approaches provide new insights into the mechanisms of segment selection and packaging in rotavirus, with broader relevance to the assembly of other segmented RNA viruses.
To define these interactions, we applied RNA crosslinking approaches capable of capturing intermolecular RNA contacts, to enable us to dissect the impact of co-transcriptional folding and to compare RNA interaction networks formed by in vitro transcription versus those formed during in situ (intra-virion) transcription. In parallel, we are developing a live-cell imaging system using reverse genetics to rescue viruses suitable for real-time transcriptional imaging, offering temporal resolution beyond that of fixed-cell methods like single-molecule FISH.
Together, these approaches provide new insights into the mechanisms of segment selection and packaging in rotavirus, with broader relevance to the assembly of other segmented RNA viruses.