Rolfsson, O, Middleton, S, Manfield, IW et al. (9 more authors) (2016) Direct Evidence for Packaging Signal-mediated Assembly of Bacteriophage MS2. Journal of Molecular Biology, 428 (2). pp. 431-448. ISSN 0022-2836
Abstract
Using cross-linking coupled to MALDI mass spectrometry and CLIP-Seq sequencing we determined the peptide and oligonucleotide sequences at the interfaces between the capsid proteins and the genomic RNA of bacteriophage MS2. The results suggest that the same coat protein (CP)-RNA and maturation protein (MP)-RNA interfaces are used in every viral particle. The portions of the viral RNA in contact with CP subunits span the genome, consistent with a large number of discrete and similar contacts within each particle. Many of these sites match previous predictions of the locations of multiple, dispersed and degenerate RNA sites with cognate CP affinity termed packaging signals (PSs). Chemical RNA footprinting was used to compare the secondary structures of protein-free genomic fragments and the RNA in the virion. Some PSs are partially present in protein-free RNA but others would need to refold from their dominant solution conformations to form the contacts identified in the virion. The RNA-binding peptides within the MP map to two sections of the N-terminal half of the protein. Comparison of MP sequences from related phages suggests a similar arrangement of RNA-binding sites, although these N-terminal regions have only limited sequence conservation. In contrast, the sequences of the C-termini are highly conserved, consistent with them encompassing pilin-binding domains required for initial contact with host cells. These results provide independent and unambiguous support for the assembly of MS2 virions via a PS-mediated mechanism involving a series of induced-fit viral protein interactions with RNA.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2015 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
Keywords: | RNA virus; assembly mechanism; RNA-protein interaction; packaging signals |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) |
Funding Information: | Funder Grant number BBSRC BB/L022095/1 Wellcome Trust 090932/Z/09/Z BBSRC BB/J00667X/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 04 Dec 2015 14:44 |
Last Modified: | 07 Aug 2018 14:12 |
Published Version: | http://dx.doi.org/10.1016/j.jmb.2015.11.014 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.jmb.2015.11.014 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:92521 |