Cargando…

Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo

The 2014–2016 Zika virus (ZIKV) epidemic in the Americas resulted in large deposits of next-generation sequencing data from clinical samples. This resource was mined to identify emerging mutations and trends in mutations as the outbreak progressed over time. Information on transmission dynamics, pre...

Descripción completa

Detalles Bibliográficos
Autores principales: Collette, Nicole M., Lao, Victoria H. I., Weilhammer, Dina R., Zingg, Barbara, Cohen, Shoshana D., Hwang, Mona, Coffey, Lark L., Grady, Sarah L., Zemla, Adam T., Borucki, Monica K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7697975/
https://www.ncbi.nlm.nih.gov/pubmed/33198111
http://dx.doi.org/10.3390/v12111295
_version_ 1783615722551771136
author Collette, Nicole M.
Lao, Victoria H. I.
Weilhammer, Dina R.
Zingg, Barbara
Cohen, Shoshana D.
Hwang, Mona
Coffey, Lark L.
Grady, Sarah L.
Zemla, Adam T.
Borucki, Monica K.
author_facet Collette, Nicole M.
Lao, Victoria H. I.
Weilhammer, Dina R.
Zingg, Barbara
Cohen, Shoshana D.
Hwang, Mona
Coffey, Lark L.
Grady, Sarah L.
Zemla, Adam T.
Borucki, Monica K.
author_sort Collette, Nicole M.
collection PubMed
description The 2014–2016 Zika virus (ZIKV) epidemic in the Americas resulted in large deposits of next-generation sequencing data from clinical samples. This resource was mined to identify emerging mutations and trends in mutations as the outbreak progressed over time. Information on transmission dynamics, prevalence, and persistence of intra-host mutants, and the position of a mutation on a protein were then used to prioritize 544 reported mutations based on their ability to impact ZIKV phenotype. Using this criteria, six mutants (representing naturally occurring mutations) were generated as synthetic infectious clones using a 2015 Puerto Rican epidemic strain PRVABC59 as the parental backbone. The phenotypes of these naturally occurring variants were examined using both cell culture and murine model systems. Mutants had distinct phenotypes, including changes in replication rate, embryo death, and decreased head size. In particular, a NS2B mutant previously detected during in vivo studies in rhesus macaques was found to cause lethal infections in adult mice, abortions in pregnant females, and increased viral genome copies in both brain tissue and blood of female mice. Additionally, mutants with changes in the region of NS3 that interfaces with NS5 during replication displayed reduced replication in the blood of adult mice. This analytical pathway, integrating both bioinformatic and wet lab experiments, provides a foundation for understanding how naturally occurring single mutations affect disease outcome and can be used to predict the of severity of future ZIKV outbreaks. To determine if naturally occurring individual mutations in the Zika virus epidemic genotype affect viral virulence or replication rate in vitro or in vivo, we generated an infectious clone representing the epidemic genotype of stain Puerto Rico, 2015. Using this clone, six mutants were created by changing nucleotides in the genome to cause one to two amino acid substitutions in the encoded proteins. The six mutants we generated represent mutations that differentiated the early epidemic genotype from genotypes that were either ancestral or that occurred later in the epidemic. We assayed each mutant for changes in growth rate, and for virulence in adult mice and pregnant mice. Three of the mutants caused catastrophic embryo effects including increased embryonic death or significant decrease in head diameter. Three other mutants that had mutations in a genome region associated with replication resulted in changes in in vitro and in vivo replication rates. These results illustrate the potential impact of individual mutations in viral phenotype.
format Online
Article
Text
id pubmed-7697975
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-76979752020-11-29 Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo Collette, Nicole M. Lao, Victoria H. I. Weilhammer, Dina R. Zingg, Barbara Cohen, Shoshana D. Hwang, Mona Coffey, Lark L. Grady, Sarah L. Zemla, Adam T. Borucki, Monica K. Viruses Article The 2014–2016 Zika virus (ZIKV) epidemic in the Americas resulted in large deposits of next-generation sequencing data from clinical samples. This resource was mined to identify emerging mutations and trends in mutations as the outbreak progressed over time. Information on transmission dynamics, prevalence, and persistence of intra-host mutants, and the position of a mutation on a protein were then used to prioritize 544 reported mutations based on their ability to impact ZIKV phenotype. Using this criteria, six mutants (representing naturally occurring mutations) were generated as synthetic infectious clones using a 2015 Puerto Rican epidemic strain PRVABC59 as the parental backbone. The phenotypes of these naturally occurring variants were examined using both cell culture and murine model systems. Mutants had distinct phenotypes, including changes in replication rate, embryo death, and decreased head size. In particular, a NS2B mutant previously detected during in vivo studies in rhesus macaques was found to cause lethal infections in adult mice, abortions in pregnant females, and increased viral genome copies in both brain tissue and blood of female mice. Additionally, mutants with changes in the region of NS3 that interfaces with NS5 during replication displayed reduced replication in the blood of adult mice. This analytical pathway, integrating both bioinformatic and wet lab experiments, provides a foundation for understanding how naturally occurring single mutations affect disease outcome and can be used to predict the of severity of future ZIKV outbreaks. To determine if naturally occurring individual mutations in the Zika virus epidemic genotype affect viral virulence or replication rate in vitro or in vivo, we generated an infectious clone representing the epidemic genotype of stain Puerto Rico, 2015. Using this clone, six mutants were created by changing nucleotides in the genome to cause one to two amino acid substitutions in the encoded proteins. The six mutants we generated represent mutations that differentiated the early epidemic genotype from genotypes that were either ancestral or that occurred later in the epidemic. We assayed each mutant for changes in growth rate, and for virulence in adult mice and pregnant mice. Three of the mutants caused catastrophic embryo effects including increased embryonic death or significant decrease in head diameter. Three other mutants that had mutations in a genome region associated with replication resulted in changes in in vitro and in vivo replication rates. These results illustrate the potential impact of individual mutations in viral phenotype. MDPI 2020-11-12 /pmc/articles/PMC7697975/ /pubmed/33198111 http://dx.doi.org/10.3390/v12111295 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Collette, Nicole M.
Lao, Victoria H. I.
Weilhammer, Dina R.
Zingg, Barbara
Cohen, Shoshana D.
Hwang, Mona
Coffey, Lark L.
Grady, Sarah L.
Zemla, Adam T.
Borucki, Monica K.
Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo
title Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo
title_full Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo
title_fullStr Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo
title_full_unstemmed Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo
title_short Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo
title_sort single amino acid mutations affect zika virus replication in vitro and virulence in vivo
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7697975/
https://www.ncbi.nlm.nih.gov/pubmed/33198111
http://dx.doi.org/10.3390/v12111295
work_keys_str_mv AT collettenicolem singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT laovictoriahi singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT weilhammerdinar singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT zinggbarbara singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT cohenshoshanad singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT hwangmona singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT coffeylarkl singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT gradysarahl singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT zemlaadamt singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo
AT boruckimonicak singleaminoacidmutationsaffectzikavirusreplicationinvitroandvirulenceinvivo