Coevolutionary constraints of Zika virus nonstructural protein 5 replication and interferon antagonism activities
Abstract
The flavivirus nonstructural protein 5 performs multiple functions during infection, including RNA replication and type I interferon signaling antagonism. Although flavivirus NS5 proteins inhibit IFN signaling through distinct mechanisms, which suggests evolutionary flexibility, the evolutionary constraints for these activities to coexist within a single protein remain to be determined. Here, we map the Zika virus NS5 STAT2 antagonism determinants and compare them with replication constraints defined by deep mutational scanning. Antagonism and replication determinants extensively overlap, and no single amino acid substitution eliminates antagonism without impairing replication. Resolving these fitness landscapes in parallel identifies specific combinations of partially functional substitutions that retain replication capacity while markedly reducing antagonism. These viruses are profoundly attenuated in human STAT2 knock-in mice. Our results uncover a fundamental evolutionary constraint linking replication and immune evasion activities in NS5, highlight that STAT2 antagonism is essential for ZIKV pathogenesis and provide new avenues for attenuated ZIKV vaccines.
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BibTeX
@article{richardson2026coevolutionary,
title = {{Coevolutionary constraints of Zika virus nonstructural protein 5 replication and interferon antagonism activities}},
author = {R. Blake Richardson and Caroline Kikawa and Amit Garg and Eva Bednarski and Madihah Salim and David J. Bacsik and Ethan C. Veit and Anna Hermacinski and Rachael Hamilton and Adolfo García-Sastre and Jesse D. Bloom and Jean K. Lim and Matthew J. Evans},
journal = {Nature Communications},
year = {2026},
month = oct,
doi = {10.1038/s41467-026-78902-1},
url = {https://www.nature.com/articles/s41467-026-78902-1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Coevolutionary constraints of Zika virus nonstructural protein 5 replication and interferon antagonism activities
AU - R. Blake Richardson
AU - Caroline Kikawa
AU - Amit Garg
AU - Eva Bednarski
AU - Madihah Salim
AU - David J. Bacsik
AU - Ethan C. Veit
AU - Anna Hermacinski
AU - Rachael Hamilton
AU - Adolfo García-Sastre
AU - Jesse D. Bloom
AU - Jean K. Lim
AU - Matthew J. Evans
PY - 2026
DA - 2026/10/28
JO - Nature Communications
DO - 10.1038/s41467-026-78902-1
UR - https://www.nature.com/articles/s41467-026-78902-1
AB - The flavivirus nonstructural protein 5 performs multiple functions during infection, including RNA replication and type I interferon signaling antagonism. Although flavivirus NS5 proteins inhibit IFN signaling through distinct mechanisms, which suggests evolutionary flexibility, the evolutionary constraints for these activities to coexist within a single protein remain to be determined. Here, we map the Zika virus NS5 STAT2 antagonism determinants and compare them with replication constraints defined by deep mutational scanning. Antagonism and replication determinants extensively overlap, and no single amino acid substitution eliminates antagonism without impairing replication. Resolving these fitness landscapes in parallel identifies specific combinations of partially functional substitutions that retain replication capacity while markedly reducing antagonism. These viruses are profoundly attenuated in human STAT2 knock-in mice. Our results uncover a fundamental evolutionary constraint linking replication and immune evasion activities in NS5, highlight that STAT2 antagonism is essential for ZIKV pathogenesis and provide new avenues for attenuated ZIKV vaccines.
SN - 2041-1723
ER -