Format

Send to

Choose Destination
Elife. 2017 Sep 18;6. pii: e27453. doi: 10.7554/eLife.27453.

Protein-mediated RNA folding governs sequence-specific interactions between rotavirus genome segments.

Author information

1
Astbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom.
2
Department of Chemistry, Center for NanoScience, Nanosystems Initiative Munich (NIM) and Center for Integrated Protein Science Munich (CiPSM), Ludwig-Maximilian University of Munich, Munich, Germany.
3
York Centre for Complex Systems Analysis, University of York, York, United Kingdom.
4
Department of Mathematics, University of York, York, United Kingdom.
5
Department of Biology, University of York, York, United Kingdom.

Abstract

Segmented RNA viruses are ubiquitous pathogens, which include influenza viruses and rotaviruses. A major challenge in understanding their assembly is the combinatorial problem of a non-random selection of a full genomic set of distinct RNAs. This process involves complex RNA-RNA and protein-RNA interactions, which are often obscured by non-specific binding at concentrations approaching in vivo assembly conditions. Here, we present direct experimental evidence of sequence-specific inter-segment interactions between rotavirus RNAs, taking place in a complex RNA- and protein-rich milieu. We show that binding of the rotavirus-encoded non-structural protein NSP2 to viral ssRNAs results in the remodeling of RNA, which is conducive to formation of stable inter-segment contacts. To identify the sites of these interactions, we have developed an RNA-RNA SELEX approach for mapping the sequences involved in inter-segment base-pairing. Our findings elucidate the molecular basis underlying inter-segment interactions in rotaviruses, paving the way for delineating similar RNA-RNA interactions that govern assembly of other segmented RNA viruses.

KEYWORDS:

Fluorescence Cross-Correlation Spectroscopy; RNA folding; RNA-RNA interactions; Rotavirus; Virus assembly; biophysics; infectious disease; microbiology; structural biology; virus

PMID:
28922109
PMCID:
PMC5621836
DOI:
10.7554/eLife.27453
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for eLife Sciences Publications, Ltd Icon for PubMed Central
Loading ...
Support Center