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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...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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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. |
format | Online Article Text |
id | pubmed-8707032 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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