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High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms

Negative-strand RNA viruses condense their genome into helical nucleocapsids that constitute essential templates for viral replication and transcription. The intrinsic flexibility of nucleocapsids usually prevents their full-length structural characterisation at high resolution. Here, we describe pu...

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Autores principales: Arragain, Benoît, Reguera, Juan, Desfosses, Ambroise, Gutsche, Irina, Schoehn, Guy, Malet, Hélène
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6365055/
https://www.ncbi.nlm.nih.gov/pubmed/30638449
http://dx.doi.org/10.7554/eLife.43075
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author Arragain, Benoît
Reguera, Juan
Desfosses, Ambroise
Gutsche, Irina
Schoehn, Guy
Malet, Hélène
author_facet Arragain, Benoît
Reguera, Juan
Desfosses, Ambroise
Gutsche, Irina
Schoehn, Guy
Malet, Hélène
author_sort Arragain, Benoît
collection PubMed
description Negative-strand RNA viruses condense their genome into helical nucleocapsids that constitute essential templates for viral replication and transcription. The intrinsic flexibility of nucleocapsids usually prevents their full-length structural characterisation at high resolution. Here, we describe purification of full-length recombinant metastable helical nucleocapsid of Hantaan virus (Hantaviridae family, Bunyavirales order) and determine its structure at 3.3 Å resolution by cryo-electron microscopy. The structure reveals the mechanisms of helical multimerisation via sub-domain exchanges between protomers and highlights nucleotide positions in a continuous positively charged groove compatible with viral genome binding. It uncovers key sites for future structure-based design of antivirals that are currently lacking to counteract life-threatening hantavirus infections. The structure also suggests a model of nucleoprotein-polymerase interaction that would enable replication and transcription solely upon local disruption of the nucleocapsid.
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spelling pubmed-63650552019-02-07 High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms Arragain, Benoît Reguera, Juan Desfosses, Ambroise Gutsche, Irina Schoehn, Guy Malet, Hélène eLife Structural Biology and Molecular Biophysics Negative-strand RNA viruses condense their genome into helical nucleocapsids that constitute essential templates for viral replication and transcription. The intrinsic flexibility of nucleocapsids usually prevents their full-length structural characterisation at high resolution. Here, we describe purification of full-length recombinant metastable helical nucleocapsid of Hantaan virus (Hantaviridae family, Bunyavirales order) and determine its structure at 3.3 Å resolution by cryo-electron microscopy. The structure reveals the mechanisms of helical multimerisation via sub-domain exchanges between protomers and highlights nucleotide positions in a continuous positively charged groove compatible with viral genome binding. It uncovers key sites for future structure-based design of antivirals that are currently lacking to counteract life-threatening hantavirus infections. The structure also suggests a model of nucleoprotein-polymerase interaction that would enable replication and transcription solely upon local disruption of the nucleocapsid. eLife Sciences Publications, Ltd 2019-01-14 /pmc/articles/PMC6365055/ /pubmed/30638449 http://dx.doi.org/10.7554/eLife.43075 Text en © 2019, Arragain et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Structural Biology and Molecular Biophysics
Arragain, Benoît
Reguera, Juan
Desfosses, Ambroise
Gutsche, Irina
Schoehn, Guy
Malet, Hélène
High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms
title High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms
title_full High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms
title_fullStr High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms
title_full_unstemmed High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms
title_short High resolution cryo-EM structure of the helical RNA-bound Hantaan virus nucleocapsid reveals its assembly mechanisms
title_sort high resolution cryo-em structure of the helical rna-bound hantaan virus nucleocapsid reveals its assembly mechanisms
topic Structural Biology and Molecular Biophysics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6365055/
https://www.ncbi.nlm.nih.gov/pubmed/30638449
http://dx.doi.org/10.7554/eLife.43075
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