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Rewriting Nature’s ssRNA Virus Assembly Manual

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JournalProceedings of the National Academy of Sciences of the United States of America
DateAccepted/In press - 3 Oct 2017
DatePublished (current) - 14 Nov 2017
Issue number46
Volume114
Number of pages6
Pages (from-to)12255-12260
Original languageEnglish

Abstract

Satellite Tobacco Necrosis Virus (STNV) is one of the smallest viruses known. Its genome encodes only its coat protein subunit relying on the polymerase of its helper virus TNV for replication. The genome has been shown to contain a cryptic set of dispersed assembly signals in the form of stem-loops that each present a minimal coat protein binding motif –A.X.X.A- in the loops. The 5′ 127 nt fragment is predicted to encompass five such Packaging Signals (PSs). We have used mutagenesis to determine the critical assembly features in this region of the STNV genome. These include the coat protein binding motif, the relative placement of PS stem-loops, their number and their folding propensity. Coat protein binding has an electrostatic contribution but assembly nucleation is dominated by folding of the RNA fragment. Replacement of the viral sequence by a synthetic equivalent with superior features creates an RNA that is a better assembly substrate than the wild-type fragment. Replacing this region in the STNV1 genome with the synthetic fragment leads to better assembly of genome-length RNA compared to the wild-type sequence. These data confirm details of the PS-mediated assembly mechanism for this virus, and paves the way for in vitro production of stable virus-like particles encapsidating non-native RNAs or other cargoes.

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    Research areas

  • Satellite tobacco necrosis virus, packaging signals, viral assembly, synthetic virology

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