Cargando…

Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene

The influenza A virus genome comprises eight negative-sense viral RNAs (vRNAs) that form individual ribonucleoprotein (RNP) complexes. In order to incorporate a complete set of each of these vRNAs, the virus uses a selective packaging mechanism that facilitates co-packaging of specific gene segments...

Descripción completa

Detalles Bibliográficos
Autores principales: Gilbertson, Brad, Zheng, Tian, Gerber, Marie, Printz-Schweigert, Anne, Ong, Chi, Marquet, Roland, Isel, Catherine, Rockman, Steven, Brown, Lorena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4997600/
https://www.ncbi.nlm.nih.gov/pubmed/27556479
http://dx.doi.org/10.3390/v8080238
_version_ 1782449810432851968
author Gilbertson, Brad
Zheng, Tian
Gerber, Marie
Printz-Schweigert, Anne
Ong, Chi
Marquet, Roland
Isel, Catherine
Rockman, Steven
Brown, Lorena
author_facet Gilbertson, Brad
Zheng, Tian
Gerber, Marie
Printz-Schweigert, Anne
Ong, Chi
Marquet, Roland
Isel, Catherine
Rockman, Steven
Brown, Lorena
author_sort Gilbertson, Brad
collection PubMed
description The influenza A virus genome comprises eight negative-sense viral RNAs (vRNAs) that form individual ribonucleoprotein (RNP) complexes. In order to incorporate a complete set of each of these vRNAs, the virus uses a selective packaging mechanism that facilitates co-packaging of specific gene segments but whose molecular basis is still not fully understood. Recently, we used a competitive transfection model where plasmids encoding the A/Puerto Rico/8/34 (PR8) and A/Udorn/307/72 (Udorn) PB1 gene segments were competed to show that the Udorn PB1 gene segment is preferentially co-packaged into progeny virions with the Udorn NA gene segment. Here we created chimeric PB1 genes combining both Udorn and PR8 PB1 sequences to further define the location within the Udorn PB1 gene that drives co-segregation of these genes and show that nucleotides 1776–2070 of the PB1 gene are crucial for preferential selection. In vitro assays examining specific interactions between Udorn NA vRNA and purified vRNAs transcribed from chimeric PB1 genes also supported the importance of this region in the PB1-NA interaction. Hence, this work identifies an association between viral genes that are co-selected during packaging. It also reveals a region potentially important in the RNP-RNP interactions within the supramolecular complex that is predicted to form prior to budding to allow one of each segment to be packaged in the viral progeny. Our study lays the foundation to understand the co-selection of specific genes, which may be critical to the emergence of new viruses with pandemic potential.
format Online
Article
Text
id pubmed-4997600
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-49976002016-08-26 Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene Gilbertson, Brad Zheng, Tian Gerber, Marie Printz-Schweigert, Anne Ong, Chi Marquet, Roland Isel, Catherine Rockman, Steven Brown, Lorena Viruses Article The influenza A virus genome comprises eight negative-sense viral RNAs (vRNAs) that form individual ribonucleoprotein (RNP) complexes. In order to incorporate a complete set of each of these vRNAs, the virus uses a selective packaging mechanism that facilitates co-packaging of specific gene segments but whose molecular basis is still not fully understood. Recently, we used a competitive transfection model where plasmids encoding the A/Puerto Rico/8/34 (PR8) and A/Udorn/307/72 (Udorn) PB1 gene segments were competed to show that the Udorn PB1 gene segment is preferentially co-packaged into progeny virions with the Udorn NA gene segment. Here we created chimeric PB1 genes combining both Udorn and PR8 PB1 sequences to further define the location within the Udorn PB1 gene that drives co-segregation of these genes and show that nucleotides 1776–2070 of the PB1 gene are crucial for preferential selection. In vitro assays examining specific interactions between Udorn NA vRNA and purified vRNAs transcribed from chimeric PB1 genes also supported the importance of this region in the PB1-NA interaction. Hence, this work identifies an association between viral genes that are co-selected during packaging. It also reveals a region potentially important in the RNP-RNP interactions within the supramolecular complex that is predicted to form prior to budding to allow one of each segment to be packaged in the viral progeny. Our study lays the foundation to understand the co-selection of specific genes, which may be critical to the emergence of new viruses with pandemic potential. MDPI 2016-08-20 /pmc/articles/PMC4997600/ /pubmed/27556479 http://dx.doi.org/10.3390/v8080238 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gilbertson, Brad
Zheng, Tian
Gerber, Marie
Printz-Schweigert, Anne
Ong, Chi
Marquet, Roland
Isel, Catherine
Rockman, Steven
Brown, Lorena
Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene
title Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene
title_full Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene
title_fullStr Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene
title_full_unstemmed Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene
title_short Influenza NA and PB1 Gene Segments Interact during the Formation of Viral Progeny: Localization of the Binding Region within the PB1 Gene
title_sort influenza na and pb1 gene segments interact during the formation of viral progeny: localization of the binding region within the pb1 gene
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4997600/
https://www.ncbi.nlm.nih.gov/pubmed/27556479
http://dx.doi.org/10.3390/v8080238
work_keys_str_mv AT gilbertsonbrad influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT zhengtian influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT gerbermarie influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT printzschweigertanne influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT ongchi influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT marquetroland influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT iselcatherine influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT rockmansteven influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene
AT brownlorena influenzanaandpb1genesegmentsinteractduringtheformationofviralprogenylocalizationofthebindingregionwithinthepb1gene