NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo.
Flaviviruses such as yellow fever, dengue or Zika viruses are responsible for significant human and veterinary diseases worldwide. These viruses contain an RNA genome, prone to mutations, which enhances their potential to emerge as pathogens. Bamaga virus (BgV) is a mosquito-borne flavivirus in the...
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2020
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ftdoajarticles:oai:doaj.org/article:59337a2a4492434c922e16d1e29d0cd4 2023-05-15T15:16:26+02:00 NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo. Agathe M G Colmant Helle Bielefeldt-Ohmann Laura J Vet Caitlin A O'Brien Richard A Bowen Airn E Hartwig Steven Davis Thisun B H Piyasena Gervais Habarugira Jessica J Harrison Jody Hobson-Peters Roy A Hall 2020-03-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0008166 https://doaj.org/article/59337a2a4492434c922e16d1e29d0cd4 EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0008166 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008166 https://doaj.org/article/59337a2a4492434c922e16d1e29d0cd4 PLoS Neglected Tropical Diseases, Vol 14, Iss 3, p e0008166 (2020) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2020 ftdoajarticles https://doi.org/10.1371/journal.pntd.0008166 2022-12-31T06:00:24Z Flaviviruses such as yellow fever, dengue or Zika viruses are responsible for significant human and veterinary diseases worldwide. These viruses contain an RNA genome, prone to mutations, which enhances their potential to emerge as pathogens. Bamaga virus (BgV) is a mosquito-borne flavivirus in the yellow fever virus group that we have previously shown to be host-restricted in vertebrates and horizontally transmissible by Culex mosquitoes. Here, we aimed to characterise BgV host-restriction and to investigate the mechanisms involved. We showed that BgV could not replicate in a wide range of vertebrate cell lines and animal species. We determined that the mechanisms involved in BgV host-restriction were independent of the type-1 interferon response and RNAse L activity. Using a BgV infectious clone and two chimeric viruses generated as hybrids between BgV and West Nile virus, we demonstrated that BgV host-restriction occurred post-cell entry. Notably, BgV host-restriction was shown to be temperature-dependent, as BgV replicated in all vertebrate cell lines at 34°C but only in a subset at 37°C. Serial passaging of BgV in Vero cells resulted in adaptive mutants capable of efficient replication at 37°C. The identified mutations resulted in amino acid substitutions in NS4A-S124F, NS4B-N244K and NS5-G2C, all occurring close to a viral protease cleavage site (NS4A/2K and NS4B/NS5). These mutations were reverse engineered into infectious clones of BgV, which revealed that NS4B-N244K and NS5-G2C were sufficient to restore BgV replication in vertebrate cells at 37°C, while NS4A-S124F further increased replication efficiency. When these mutant viruses were injected into immunocompetent mice, alongside BgV and West Nile virus chimeras, infection and neurovirulence were enhanced as determined by clinical scores, seroconversion, micro-neutralisation, viremia, histopathology and immunohistochemistry, confirming the involvement of these residues in the attenuation of BgV. Our studies identify a new mechanism of host-restriction ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 14 3 e0008166 |
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Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Agathe M G Colmant Helle Bielefeldt-Ohmann Laura J Vet Caitlin A O'Brien Richard A Bowen Airn E Hartwig Steven Davis Thisun B H Piyasena Gervais Habarugira Jessica J Harrison Jody Hobson-Peters Roy A Hall NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Flaviviruses such as yellow fever, dengue or Zika viruses are responsible for significant human and veterinary diseases worldwide. These viruses contain an RNA genome, prone to mutations, which enhances their potential to emerge as pathogens. Bamaga virus (BgV) is a mosquito-borne flavivirus in the yellow fever virus group that we have previously shown to be host-restricted in vertebrates and horizontally transmissible by Culex mosquitoes. Here, we aimed to characterise BgV host-restriction and to investigate the mechanisms involved. We showed that BgV could not replicate in a wide range of vertebrate cell lines and animal species. We determined that the mechanisms involved in BgV host-restriction were independent of the type-1 interferon response and RNAse L activity. Using a BgV infectious clone and two chimeric viruses generated as hybrids between BgV and West Nile virus, we demonstrated that BgV host-restriction occurred post-cell entry. Notably, BgV host-restriction was shown to be temperature-dependent, as BgV replicated in all vertebrate cell lines at 34°C but only in a subset at 37°C. Serial passaging of BgV in Vero cells resulted in adaptive mutants capable of efficient replication at 37°C. The identified mutations resulted in amino acid substitutions in NS4A-S124F, NS4B-N244K and NS5-G2C, all occurring close to a viral protease cleavage site (NS4A/2K and NS4B/NS5). These mutations were reverse engineered into infectious clones of BgV, which revealed that NS4B-N244K and NS5-G2C were sufficient to restore BgV replication in vertebrate cells at 37°C, while NS4A-S124F further increased replication efficiency. When these mutant viruses were injected into immunocompetent mice, alongside BgV and West Nile virus chimeras, infection and neurovirulence were enhanced as determined by clinical scores, seroconversion, micro-neutralisation, viremia, histopathology and immunohistochemistry, confirming the involvement of these residues in the attenuation of BgV. Our studies identify a new mechanism of host-restriction ... |
format |
Article in Journal/Newspaper |
author |
Agathe M G Colmant Helle Bielefeldt-Ohmann Laura J Vet Caitlin A O'Brien Richard A Bowen Airn E Hartwig Steven Davis Thisun B H Piyasena Gervais Habarugira Jessica J Harrison Jody Hobson-Peters Roy A Hall |
author_facet |
Agathe M G Colmant Helle Bielefeldt-Ohmann Laura J Vet Caitlin A O'Brien Richard A Bowen Airn E Hartwig Steven Davis Thisun B H Piyasena Gervais Habarugira Jessica J Harrison Jody Hobson-Peters Roy A Hall |
author_sort |
Agathe M G Colmant |
title |
NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo. |
title_short |
NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo. |
title_full |
NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo. |
title_fullStr |
NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo. |
title_full_unstemmed |
NS4/5 mutations enhance flavivirus Bamaga virus infectivity and pathogenicity in vitro and in vivo. |
title_sort |
ns4/5 mutations enhance flavivirus bamaga virus infectivity and pathogenicity in vitro and in vivo. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2020 |
url |
https://doi.org/10.1371/journal.pntd.0008166 https://doaj.org/article/59337a2a4492434c922e16d1e29d0cd4 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 14, Iss 3, p e0008166 (2020) |
op_relation |
https://doi.org/10.1371/journal.pntd.0008166 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008166 https://doaj.org/article/59337a2a4492434c922e16d1e29d0cd4 |
op_doi |
https://doi.org/10.1371/journal.pntd.0008166 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
14 |
container_issue |
3 |
container_start_page |
e0008166 |
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1766346732526895104 |