Evidence that avian reovirus ?NS is an RNA chaperone: implications for genome segment assortment.
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ABSTRACT: Reoviruses are important human, animal and plant pathogens having 10-12 segments of double-stranded genomic RNA. The mechanisms controlling the assortment and packaging of genomic segments in these viruses, remain poorly understood. RNA-protein and RNA-RNA interactions between viral genomic segment precursors have been implicated in the process. While non-structural viral RNA-binding proteins, such as avian reovirus ?NS, are essential for virus replication, the mechanism by which they assist packaging is unclear. Here we demonstrate that ?NS assembles into stable elongated hexamers in vitro, which bind single-stranded nucleic acids with high affinity, but little sequence specificity. Using ensemble and single molecule fluorescence spectroscopy, we show that ?NS also binds to a partially double-stranded RNA, resulting in gradual helix unwinding. The hexamer can bind multiple RNA molecules and exhibits strand-annealing activity, thus mediating conversion of metastable, intramolecular stem-loops into more stable heteroduplexes. We demonstrate that the ARV ?NS acts as an RNA chaperone facilitating specific RNA-RNA interactions between genomic precursors during segment assortment and packaging.
SUBMITTER: Borodavka A
PROVIDER: S-EPMC4538827 | biostudies-other | 2015 Aug
REPOSITORIES: biostudies-other
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