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Article

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

1
Biosciences and Biotechnology Division, Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA
2
Biology Department, Los Positas Community College, Livermore, CA 94551, USA
3
Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA
4
Applied Physics Laboratory, Johns Hopkins University, Laurel, MD 20723, USA
5
Computations Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94550, USA
*
Author to whom correspondence should be addressed.
Viruses 2020, 12(11), 1295; https://doi.org/10.3390/v12111295
Received: 9 October 2020 / Revised: 9 November 2020 / Accepted: 10 November 2020 / Published: 12 November 2020
(This article belongs to the Section Animal Viruses)
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. View Full-Text
Keywords: Zika virus; mutation; variant; A129 mouse model; microcephaly; infectious clone; quasispecies Zika virus; mutation; variant; A129 mouse model; microcephaly; infectious clone; quasispecies
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MDPI and ACS Style

Collette, N.M.; Lao, V.H.I.; Weilhammer, D.R.; Zingg, B.; Cohen, S.D.; Hwang, M.; Coffey, L.L.; Grady, S.L.; Zemla, A.T.; Borucki, M.K. Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo. Viruses 2020, 12, 1295. https://doi.org/10.3390/v12111295

AMA Style

Collette NM, Lao VHI, Weilhammer DR, Zingg B, Cohen SD, Hwang M, Coffey LL, Grady SL, Zemla AT, Borucki MK. Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo. Viruses. 2020; 12(11):1295. https://doi.org/10.3390/v12111295

Chicago/Turabian Style

Collette, Nicole M., Victoria H.I. Lao, Dina R. Weilhammer, Barbara Zingg, Shoshana D. Cohen, Mona Hwang, Lark L. Coffey, Sarah L. Grady, Adam T. Zemla, and Monica K. Borucki 2020. "Single Amino Acid Mutations Affect Zika Virus Replication In Vitro and Virulence In Vivo" Viruses 12, no. 11: 1295. https://doi.org/10.3390/v12111295

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