Point mutations in the West Nile virus (Flaviviridae; Flavivirus) RNA-dependent RNA polymerase alter viral fitness in a host-dependent manner in vitro and in vivo

Greta A. Van Slyke, Alexander T. Ciota, Graham G. Willsey, Joachim Jaeger, Pei-Yong Shi, Laura D. Kramer

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The West Nile virus (WNV) genome contains a single RNA-dependent RNA polymerase (RdRp) gene, which is responsible for replication of the viral genome and, as such, is an important target for antiviral therapy. Viral RdRps are known to lack proofreading capabilities and as a result viruses such as WNV exist as a mixture of viral genotypes within an infection, enabling the virus to readily emerge and adapt to new host environments. To test the consequences of subtle structural alterations remote from the RdRp active-site, the following single point mutations were engineered in the WNV NS5 RdRp coding region: T363N, A365N, and T537I; these mutations were selected in an effort to stabilize the secondary structural elements near the rNTP binding pocket of the RdRp. Mutant viruses were tested in vitro on Vero, C6/36, Culex tarsalis and DF-1 cell types and in vivo in one day old chickens and Culex pipiens mosquitoes. Plaque morphology was affected by each mutation and growth and RNA replication kinetics were altered as well. Our results demonstrate that subtle alteration of the RdRp protein away from the active site can have a significant overall biological effect on WNV fitness, and that this effect can be host-dependent.

Original languageEnglish (US)
Pages (from-to)18-24
Number of pages7
JournalVirology
Volume427
Issue number1
DOIs
StatePublished - May 25 2012
Externally publishedYes

Fingerprint

Flaviviridae
RNA Replicase
Flavivirus
West Nile virus
Point Mutation
Culex
Catalytic Domain
Viruses
Mutation
Viral Genome
Virus Diseases
Culicidae
Antiviral Agents
Chickens
Genotype
In Vitro Techniques
Genome
RNA
Growth
Genes

Keywords

  • Arbovirus
  • Flavivirus
  • Mutagenesis
  • RNA replication
  • RNA viruses

ASJC Scopus subject areas

  • Virology

Cite this

Point mutations in the West Nile virus (Flaviviridae; Flavivirus) RNA-dependent RNA polymerase alter viral fitness in a host-dependent manner in vitro and in vivo. / Van Slyke, Greta A.; Ciota, Alexander T.; Willsey, Graham G.; Jaeger, Joachim; Shi, Pei-Yong; Kramer, Laura D.

In: Virology, Vol. 427, No. 1, 25.05.2012, p. 18-24.

Research output: Contribution to journalArticle

Van Slyke, Greta A. ; Ciota, Alexander T. ; Willsey, Graham G. ; Jaeger, Joachim ; Shi, Pei-Yong ; Kramer, Laura D. / Point mutations in the West Nile virus (Flaviviridae; Flavivirus) RNA-dependent RNA polymerase alter viral fitness in a host-dependent manner in vitro and in vivo. In: Virology. 2012 ; Vol. 427, No. 1. pp. 18-24.
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