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Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus

Highly pathogenic avian influenza viruses (HPAIVs) with H5 and H7 hemagglutinin (HA) subtypes are derived from their low pathogenic counterparts following the acquisition of multiple basic amino acids in their HA cleavage site. It has been suggested that consecutive adenine residues and a stem-loop...

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Autores principales: Kida, Yurie, Okuya, Kosuke, Saito, Takeshi, Yamagishi, Junya, Ohnuma, Aiko, Hattori, Takanari, Miyamoto, Hiroko, Manzoor, Rashid, Yoshida, Reiko, Nao, Naganori, Kajihara, Masahiro, Watanabe, Tokiko, Takada, Ayato
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8707032/
https://www.ncbi.nlm.nih.gov/pubmed/34959552
http://dx.doi.org/10.3390/pathogens10121597
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author Kida, Yurie
Okuya, Kosuke
Saito, Takeshi
Yamagishi, Junya
Ohnuma, Aiko
Hattori, Takanari
Miyamoto, Hiroko
Manzoor, Rashid
Yoshida, Reiko
Nao, Naganori
Kajihara, Masahiro
Watanabe, Tokiko
Takada, Ayato
author_facet Kida, Yurie
Okuya, Kosuke
Saito, Takeshi
Yamagishi, Junya
Ohnuma, Aiko
Hattori, Takanari
Miyamoto, Hiroko
Manzoor, Rashid
Yoshida, Reiko
Nao, Naganori
Kajihara, Masahiro
Watanabe, Tokiko
Takada, Ayato
author_sort Kida, Yurie
collection PubMed
description Highly pathogenic avian influenza viruses (HPAIVs) with H5 and H7 hemagglutinin (HA) subtypes are derived from their low pathogenic counterparts following the acquisition of multiple basic amino acids in their HA cleavage site. It has been suggested that consecutive adenine residues and a stem-loop structure in the viral RNA region that encodes the cleavage site are essential for the acquisition of the polybasic cleavage site. By using a reporter assay to detect non-templated nucleotide insertions, we found that insertions more frequently occurred in the RNA region (29 nucleotide-length) encoding the cleavage site of an H5 HA gene that was predicted to have a stem-loop structure containing consecutive adenines than in a mutated corresponding RNA region that had a disrupted loop structure with fewer adenines. In virus particles generated by using reverse genetics, nucleotide insertions that created additional codons for basic amino acids were found in the RNA region encoding the cleavage site of an H5 HA gene but not in the mutated RNA region. We confirmed the presence of virus clones with the ability to replicate without trypsin in a plaque assay and to cause lethal infection in chicks. These results demonstrate that the stem-loop structure containing consecutive adenines in HA genes is a key molecular determinant for the emergence of H5 HPAIVs.
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spelling pubmed-87070322021-12-25 Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus Kida, Yurie Okuya, Kosuke Saito, Takeshi Yamagishi, Junya Ohnuma, Aiko Hattori, Takanari Miyamoto, Hiroko Manzoor, Rashid Yoshida, Reiko Nao, Naganori Kajihara, Masahiro Watanabe, Tokiko Takada, Ayato Pathogens Article Highly pathogenic avian influenza viruses (HPAIVs) with H5 and H7 hemagglutinin (HA) subtypes are derived from their low pathogenic counterparts following the acquisition of multiple basic amino acids in their HA cleavage site. It has been suggested that consecutive adenine residues and a stem-loop structure in the viral RNA region that encodes the cleavage site are essential for the acquisition of the polybasic cleavage site. By using a reporter assay to detect non-templated nucleotide insertions, we found that insertions more frequently occurred in the RNA region (29 nucleotide-length) encoding the cleavage site of an H5 HA gene that was predicted to have a stem-loop structure containing consecutive adenines than in a mutated corresponding RNA region that had a disrupted loop structure with fewer adenines. In virus particles generated by using reverse genetics, nucleotide insertions that created additional codons for basic amino acids were found in the RNA region encoding the cleavage site of an H5 HA gene but not in the mutated RNA region. We confirmed the presence of virus clones with the ability to replicate without trypsin in a plaque assay and to cause lethal infection in chicks. These results demonstrate that the stem-loop structure containing consecutive adenines in HA genes is a key molecular determinant for the emergence of H5 HPAIVs. MDPI 2021-12-09 /pmc/articles/PMC8707032/ /pubmed/34959552 http://dx.doi.org/10.3390/pathogens10121597 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kida, Yurie
Okuya, Kosuke
Saito, Takeshi
Yamagishi, Junya
Ohnuma, Aiko
Hattori, Takanari
Miyamoto, Hiroko
Manzoor, Rashid
Yoshida, Reiko
Nao, Naganori
Kajihara, Masahiro
Watanabe, Tokiko
Takada, Ayato
Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus
title Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus
title_full Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus
title_fullStr Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus
title_full_unstemmed Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus
title_short Structural Requirements in the Hemagglutinin Cleavage Site-Coding RNA Region for the Generation of Highly Pathogenic Avian Influenza Virus
title_sort structural requirements in the hemagglutinin cleavage site-coding rna region for the generation of highly pathogenic avian influenza virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8707032/
https://www.ncbi.nlm.nih.gov/pubmed/34959552
http://dx.doi.org/10.3390/pathogens10121597
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