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Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus

We have previously described the efficient homologous recombination system between 5′ subgenomic RNA3a (sgRNA3a) and genomic RNA3 of Brome mosaic virus (BMV) in barley protoplasts (Sztuba-Solińska et al., 2011a). Here, we demonstrated that sequence alterations in the coat protein (CP)-binding cis-ac...

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Autores principales: Sztuba-Solińska, Joanna, Fanning, Sean W., Horn, James R., Bujarski, Jozef J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114393/
https://www.ncbi.nlm.nih.gov/pubmed/23079110
http://dx.doi.org/10.1016/j.virusres.2012.10.001
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author Sztuba-Solińska, Joanna
Fanning, Sean W.
Horn, James R.
Bujarski, Jozef J.
author_facet Sztuba-Solińska, Joanna
Fanning, Sean W.
Horn, James R.
Bujarski, Jozef J.
author_sort Sztuba-Solińska, Joanna
collection PubMed
description We have previously described the efficient homologous recombination system between 5′ subgenomic RNA3a (sgRNA3a) and genomic RNA3 of Brome mosaic virus (BMV) in barley protoplasts (Sztuba-Solińska et al., 2011a). Here, we demonstrated that sequence alterations in the coat protein (CP)-binding cis-acting RNA motifs, the Bbox region (in the intercistronic RNA3 sequence) and the RNA3 packaging element (PE, in the movement protein ORF), reduced crossover frequencies in protoplasts. Additionally, the modification of Bbox-like element in the 5′ UTR region strongly debilitated crossovers. Along the lines of these observations, RNA3 mutants not expressing CP or expressing mutated CPs also reduced recombination. A series of reciprocal transfections demonstrated a functional crosstalk between the Bbox and PE elements. Altogether, our data imply the role of CP in sgRNA3a-directed recombination by either facilitating the interaction of the RNA substrates and/or by creating roadblocks for the viral replicase.
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spelling pubmed-71143932020-04-02 Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus Sztuba-Solińska, Joanna Fanning, Sean W. Horn, James R. Bujarski, Jozef J. Virus Res Article We have previously described the efficient homologous recombination system between 5′ subgenomic RNA3a (sgRNA3a) and genomic RNA3 of Brome mosaic virus (BMV) in barley protoplasts (Sztuba-Solińska et al., 2011a). Here, we demonstrated that sequence alterations in the coat protein (CP)-binding cis-acting RNA motifs, the Bbox region (in the intercistronic RNA3 sequence) and the RNA3 packaging element (PE, in the movement protein ORF), reduced crossover frequencies in protoplasts. Additionally, the modification of Bbox-like element in the 5′ UTR region strongly debilitated crossovers. Along the lines of these observations, RNA3 mutants not expressing CP or expressing mutated CPs also reduced recombination. A series of reciprocal transfections demonstrated a functional crosstalk between the Bbox and PE elements. Altogether, our data imply the role of CP in sgRNA3a-directed recombination by either facilitating the interaction of the RNA substrates and/or by creating roadblocks for the viral replicase. Elsevier B.V. 2012-12 2012-10-16 /pmc/articles/PMC7114393/ /pubmed/23079110 http://dx.doi.org/10.1016/j.virusres.2012.10.001 Text en Copyright © 2012 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Sztuba-Solińska, Joanna
Fanning, Sean W.
Horn, James R.
Bujarski, Jozef J.
Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus
title Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus
title_full Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus
title_fullStr Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus
title_full_unstemmed Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus
title_short Mutations in the coat protein-binding cis-acting RNA motifs debilitate RNA recombination of Brome mosaic virus
title_sort mutations in the coat protein-binding cis-acting rna motifs debilitate rna recombination of brome mosaic virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114393/
https://www.ncbi.nlm.nih.gov/pubmed/23079110
http://dx.doi.org/10.1016/j.virusres.2012.10.001
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