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Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design
Mosquito-transmitted chikungunya virus (CHIKV) is an arthritogenic alphavirus of the Togaviridae family responsible for frequent outbreaks of arthritic disease in humans. Capsid protein, a structural protein encoded by the CHIKV RNA genome, is able to translocate to the host cell nucleolus. In encep...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5358915/ https://www.ncbi.nlm.nih.gov/pubmed/28223458 http://dx.doi.org/10.1128/mBio.01970-16 |
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author | Taylor, Adam Liu, Xiang Zaid, Ali Goh, Lucas Y. H. Hobson-Peters, Jody Hall, Roy A. Merits, Andres Mahalingam, Suresh |
author_facet | Taylor, Adam Liu, Xiang Zaid, Ali Goh, Lucas Y. H. Hobson-Peters, Jody Hall, Roy A. Merits, Andres Mahalingam, Suresh |
author_sort | Taylor, Adam |
collection | PubMed |
description | Mosquito-transmitted chikungunya virus (CHIKV) is an arthritogenic alphavirus of the Togaviridae family responsible for frequent outbreaks of arthritic disease in humans. Capsid protein, a structural protein encoded by the CHIKV RNA genome, is able to translocate to the host cell nucleolus. In encephalitic alphaviruses, nuclear translocation induces host cell transcriptional shutoff; however, the role of capsid protein nucleolar localization in arthritogenic alphaviruses remains unclear. Using recombinant enhanced green fluorescent protein (EGFP)-tagged expression constructs and CHIKV infectious clones, we describe a nucleolar localization sequence (NoLS) in the N-terminal region of capsid protein, previously uncharacterized in CHIKV. Mutation of the NoLS by site-directed mutagenesis reduced efficiency of nuclear import of CHIKV capsid protein. In the virus, mutation of the capsid protein NoLS (CHIKV-NoLS) attenuated replication in mammalian and mosquito cells, producing a small-plaque phenotype. Attenuation of CHIKV-NoLS is likely due to disruption of the viral replication cycle downstream of viral RNA synthesis. In mice, CHIKV-NoLS infection caused no disease signs compared to wild-type CHIKV (CHIKV-WT)-infected mice; lack of disease signs correlated with significantly reduced viremia and decreased expression of proinflammatory factors. Mice immunized with CHIKV-NoLS, challenged with CHIKV-WT at 30 days postimmunization, develop no disease signs and no detectable viremia. Serum from CHIKV-NoLS-immunized mice is able to efficiently neutralize CHIKV infection in vitro. Additionally, CHIKV-NoLS-immunized mice challenged with the related alphavirus Ross River virus showed reduced early and peak viremia postchallenge, indicating a cross-protective effect. The high degree of CHIKV-NoLS attenuation may improve CHIKV antiviral and rational vaccine design. |
format | Online Article Text |
id | pubmed-5358915 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-53589152017-03-24 Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design Taylor, Adam Liu, Xiang Zaid, Ali Goh, Lucas Y. H. Hobson-Peters, Jody Hall, Roy A. Merits, Andres Mahalingam, Suresh mBio Research Article Mosquito-transmitted chikungunya virus (CHIKV) is an arthritogenic alphavirus of the Togaviridae family responsible for frequent outbreaks of arthritic disease in humans. Capsid protein, a structural protein encoded by the CHIKV RNA genome, is able to translocate to the host cell nucleolus. In encephalitic alphaviruses, nuclear translocation induces host cell transcriptional shutoff; however, the role of capsid protein nucleolar localization in arthritogenic alphaviruses remains unclear. Using recombinant enhanced green fluorescent protein (EGFP)-tagged expression constructs and CHIKV infectious clones, we describe a nucleolar localization sequence (NoLS) in the N-terminal region of capsid protein, previously uncharacterized in CHIKV. Mutation of the NoLS by site-directed mutagenesis reduced efficiency of nuclear import of CHIKV capsid protein. In the virus, mutation of the capsid protein NoLS (CHIKV-NoLS) attenuated replication in mammalian and mosquito cells, producing a small-plaque phenotype. Attenuation of CHIKV-NoLS is likely due to disruption of the viral replication cycle downstream of viral RNA synthesis. In mice, CHIKV-NoLS infection caused no disease signs compared to wild-type CHIKV (CHIKV-WT)-infected mice; lack of disease signs correlated with significantly reduced viremia and decreased expression of proinflammatory factors. Mice immunized with CHIKV-NoLS, challenged with CHIKV-WT at 30 days postimmunization, develop no disease signs and no detectable viremia. Serum from CHIKV-NoLS-immunized mice is able to efficiently neutralize CHIKV infection in vitro. Additionally, CHIKV-NoLS-immunized mice challenged with the related alphavirus Ross River virus showed reduced early and peak viremia postchallenge, indicating a cross-protective effect. The high degree of CHIKV-NoLS attenuation may improve CHIKV antiviral and rational vaccine design. American Society for Microbiology 2017-02-21 /pmc/articles/PMC5358915/ /pubmed/28223458 http://dx.doi.org/10.1128/mBio.01970-16 Text en Copyright © 2017 Taylor et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (http://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Article Taylor, Adam Liu, Xiang Zaid, Ali Goh, Lucas Y. H. Hobson-Peters, Jody Hall, Roy A. Merits, Andres Mahalingam, Suresh Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design |
title | Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design |
title_full | Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design |
title_fullStr | Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design |
title_full_unstemmed | Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design |
title_short | Mutation of the N-Terminal Region of Chikungunya Virus Capsid Protein: Implications for Vaccine Design |
title_sort | mutation of the n-terminal region of chikungunya virus capsid protein: implications for vaccine design |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5358915/ https://www.ncbi.nlm.nih.gov/pubmed/28223458 http://dx.doi.org/10.1128/mBio.01970-16 |
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