Cargando…

Monomeric Nucleoprotein of Influenza A Virus

Isolated influenza A virus nucleoprotein exists in an equilibrium between monomers and trimers. Samples containing only monomers or only trimers can be stabilized by respectively low and high salt. The trimers bind RNA with high affinity but remain trimmers, whereas the monomers polymerise onto RNA...

Descripción completa

Detalles Bibliográficos
Autores principales: Chenavas, Sylvie, Estrozi, Leandro F., Slama-Schwok, Anny, Delmas, Bernard, Di Primo, Carmelo, Baudin, Florence, Li, Xinping, Crépin, Thibaut, Ruigrok, Rob W. H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3610751/
https://www.ncbi.nlm.nih.gov/pubmed/23555270
http://dx.doi.org/10.1371/journal.ppat.1003275
_version_ 1782264501146484736
author Chenavas, Sylvie
Estrozi, Leandro F.
Slama-Schwok, Anny
Delmas, Bernard
Di Primo, Carmelo
Baudin, Florence
Li, Xinping
Crépin, Thibaut
Ruigrok, Rob W. H.
author_facet Chenavas, Sylvie
Estrozi, Leandro F.
Slama-Schwok, Anny
Delmas, Bernard
Di Primo, Carmelo
Baudin, Florence
Li, Xinping
Crépin, Thibaut
Ruigrok, Rob W. H.
author_sort Chenavas, Sylvie
collection PubMed
description Isolated influenza A virus nucleoprotein exists in an equilibrium between monomers and trimers. Samples containing only monomers or only trimers can be stabilized by respectively low and high salt. The trimers bind RNA with high affinity but remain trimmers, whereas the monomers polymerise onto RNA forming nucleoprotein-RNA complexes. When wild type (wt) nucleoprotein is crystallized, it forms trimers, whether one starts with monomers or trimers. We therefore crystallized the obligate monomeric R416A mutant nucleoprotein and observed how the domain exchange loop that leads over to a neighbouring protomer in the trimer structure interacts with equivalent sites on the mutant monomer surface, avoiding polymerisation. The C-terminus of the monomer is bound to the side of the RNA binding surface, lowering its positive charge. Biophysical characterization of the mutant and wild type monomeric proteins gives the same results, suggesting that the exchange domain is folded in the same way for the wild type protein. In a search for how monomeric wt nucleoprotein may be stabilized in the infected cell we determined the phosphorylation sites on nucleoprotein isolated from virus particles. We found that serine 165 was phosphorylated and conserved in all influenza A and B viruses. The S165D mutant that mimics phosphorylation is monomeric and displays a lowered affinity for RNA compared with wt monomeric NP. This suggests that phosphorylation may regulate the polymerisation state and RNA binding of nucleoprotein in the infected cell. The monomer structure could be used for finding new anti influenza drugs because compounds that stabilize the monomer may slow down viral infection.
format Online
Article
Text
id pubmed-3610751
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-36107512013-04-03 Monomeric Nucleoprotein of Influenza A Virus Chenavas, Sylvie Estrozi, Leandro F. Slama-Schwok, Anny Delmas, Bernard Di Primo, Carmelo Baudin, Florence Li, Xinping Crépin, Thibaut Ruigrok, Rob W. H. PLoS Pathog Research Article Isolated influenza A virus nucleoprotein exists in an equilibrium between monomers and trimers. Samples containing only monomers or only trimers can be stabilized by respectively low and high salt. The trimers bind RNA with high affinity but remain trimmers, whereas the monomers polymerise onto RNA forming nucleoprotein-RNA complexes. When wild type (wt) nucleoprotein is crystallized, it forms trimers, whether one starts with monomers or trimers. We therefore crystallized the obligate monomeric R416A mutant nucleoprotein and observed how the domain exchange loop that leads over to a neighbouring protomer in the trimer structure interacts with equivalent sites on the mutant monomer surface, avoiding polymerisation. The C-terminus of the monomer is bound to the side of the RNA binding surface, lowering its positive charge. Biophysical characterization of the mutant and wild type monomeric proteins gives the same results, suggesting that the exchange domain is folded in the same way for the wild type protein. In a search for how monomeric wt nucleoprotein may be stabilized in the infected cell we determined the phosphorylation sites on nucleoprotein isolated from virus particles. We found that serine 165 was phosphorylated and conserved in all influenza A and B viruses. The S165D mutant that mimics phosphorylation is monomeric and displays a lowered affinity for RNA compared with wt monomeric NP. This suggests that phosphorylation may regulate the polymerisation state and RNA binding of nucleoprotein in the infected cell. The monomer structure could be used for finding new anti influenza drugs because compounds that stabilize the monomer may slow down viral infection. Public Library of Science 2013-03-28 /pmc/articles/PMC3610751/ /pubmed/23555270 http://dx.doi.org/10.1371/journal.ppat.1003275 Text en © 2013 Chenavas et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chenavas, Sylvie
Estrozi, Leandro F.
Slama-Schwok, Anny
Delmas, Bernard
Di Primo, Carmelo
Baudin, Florence
Li, Xinping
Crépin, Thibaut
Ruigrok, Rob W. H.
Monomeric Nucleoprotein of Influenza A Virus
title Monomeric Nucleoprotein of Influenza A Virus
title_full Monomeric Nucleoprotein of Influenza A Virus
title_fullStr Monomeric Nucleoprotein of Influenza A Virus
title_full_unstemmed Monomeric Nucleoprotein of Influenza A Virus
title_short Monomeric Nucleoprotein of Influenza A Virus
title_sort monomeric nucleoprotein of influenza a virus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3610751/
https://www.ncbi.nlm.nih.gov/pubmed/23555270
http://dx.doi.org/10.1371/journal.ppat.1003275
work_keys_str_mv AT chenavassylvie monomericnucleoproteinofinfluenzaavirus
AT estrozileandrof monomericnucleoproteinofinfluenzaavirus
AT slamaschwokanny monomericnucleoproteinofinfluenzaavirus
AT delmasbernard monomericnucleoproteinofinfluenzaavirus
AT diprimocarmelo monomericnucleoproteinofinfluenzaavirus
AT baudinflorence monomericnucleoproteinofinfluenzaavirus
AT lixinping monomericnucleoproteinofinfluenzaavirus
AT crepinthibaut monomericnucleoproteinofinfluenzaavirus
AT ruigrokrobwh monomericnucleoproteinofinfluenzaavirus