Mutations in West Nile virus nonstructural proteins that facilitate replicon persistence in vitro attenuate virus replication in vitro and in vivo

Shannan L. Rossi; Rafik Fayzulin; Nathan Dewsbury; Nigel Bourne; Peter W. Mason

doi:10.1016/j.virol.2007.02.009

Mutations in West Nile virus nonstructural proteins that facilitate replicon persistence in vitro attenuate virus replication in vitro and in vivo

Shannan L. Rossi, Rafik Fayzulin, Nathan Dewsbury, Nigel Bourne, Peter W. Mason

Research output: Contribution to journal › Article › peer-review

38 Scopus citations

Abstract

West Nile virus (WNV) infections in vertebrates are generally acute but persistent infections have been observed. To investigate the ability of WNV to produce persistent infections, we forced subgenomic WNV replicons to replicate within a cell without causing cell death. Detailed analyses of these cell-adapted genomes revealed mutations within the nonstructural protein genes NS2A (D73H, M108K), NS3 (117Kins), NS4B (E249G) and NS5 (P528H). WNV replicons and WNVs harboring a subset of NS2A or NS3 mutations showed a reduction in genome replication, a reduction in antigen accumulation, a decrease in cytopathic effect, an increased ability to persist in cell culture and/or attenuation in vivo. Taken together, these data indicate that WNV with a defect in replication and an increased potential to persist within the host cell can be generated by point mutations at multiple independent loci, suggesting that persistent viruses could arise in nature.

Original language	English (US)
Pages (from-to)	184-195
Number of pages	12
Journal	Virology
Volume	364
Issue number	1
DOIs	https://doi.org/10.1016/j.virol.2007.02.009
State	Published - Jul 20 2007

Keywords

Attenuation
Flavivirus
Innate immune response
Interferon
Nonstructural gene
Persistence
Replication
Replicon
West Nile virus

ASJC Scopus subject areas

Virology

Access to Document

10.1016/j.virol.2007.02.009

Cite this

@article{497f51ba0efc4063b72fb5f52fc41e0f,

title = "Mutations in West Nile virus nonstructural proteins that facilitate replicon persistence in vitro attenuate virus replication in vitro and in vivo",

abstract = "West Nile virus (WNV) infections in vertebrates are generally acute but persistent infections have been observed. To investigate the ability of WNV to produce persistent infections, we forced subgenomic WNV replicons to replicate within a cell without causing cell death. Detailed analyses of these cell-adapted genomes revealed mutations within the nonstructural protein genes NS2A (D73H, M108K), NS3 (117Kins), NS4B (E249G) and NS5 (P528H). WNV replicons and WNVs harboring a subset of NS2A or NS3 mutations showed a reduction in genome replication, a reduction in antigen accumulation, a decrease in cytopathic effect, an increased ability to persist in cell culture and/or attenuation in vivo. Taken together, these data indicate that WNV with a defect in replication and an increased potential to persist within the host cell can be generated by point mutations at multiple independent loci, suggesting that persistent viruses could arise in nature.",

keywords = "Attenuation, Flavivirus, Innate immune response, Interferon, Nonstructural gene, Persistence, Replication, Replicon, West Nile virus",

author = "Rossi, {Shannan L.} and Rafik Fayzulin and Nathan Dewsbury and Nigel Bourne and Mason, {Peter W.}",

note = "Funding Information: We thank J. Durbin of Ohio State University for providing the STAT1 −/− MEF cell line, I. Frolov for providing the MEF RNase L −/− cell line and BAC plasmid and R. Suzuki for cloning the WNV infectious clone cDNA into a derivative of this BAC plasmid. We also thank F.D. Gilfoy for her help in generating replicon-bearing cells used in this study. We are grateful for the technical assistance of E. Infante and T. Ethridge. We are very grateful for F. Scholle and N. Vasilakis for their critical review of the manuscript. S.L.R. was supported by the James McLaughlin fellowship and a NIH training grant (T32 060549-01). This work was supported by a grant from NIAID to P.W.M. through the Western Regional Center of Excellence for Biodefense and Emerging Infectious Disease Research (NIH grant number U54 AI057156). ",

year = "2007",

month = jul,

day = "20",

doi = "10.1016/j.virol.2007.02.009",

language = "English (US)",

volume = "364",

pages = "184--195",

journal = "Virology",

issn = "0042-6822",

publisher = "Academic Press Inc.",

number = "1",

}

TY - JOUR

T1 - Mutations in West Nile virus nonstructural proteins that facilitate replicon persistence in vitro attenuate virus replication in vitro and in vivo

AU - Rossi, Shannan L.

AU - Fayzulin, Rafik

AU - Dewsbury, Nathan

AU - Bourne, Nigel

AU - Mason, Peter W.

N1 - Funding Information: We thank J. Durbin of Ohio State University for providing the STAT1 −/− MEF cell line, I. Frolov for providing the MEF RNase L −/− cell line and BAC plasmid and R. Suzuki for cloning the WNV infectious clone cDNA into a derivative of this BAC plasmid. We also thank F.D. Gilfoy for her help in generating replicon-bearing cells used in this study. We are grateful for the technical assistance of E. Infante and T. Ethridge. We are very grateful for F. Scholle and N. Vasilakis for their critical review of the manuscript. S.L.R. was supported by the James McLaughlin fellowship and a NIH training grant (T32 060549-01). This work was supported by a grant from NIAID to P.W.M. through the Western Regional Center of Excellence for Biodefense and Emerging Infectious Disease Research (NIH grant number U54 AI057156).

PY - 2007/7/20

Y1 - 2007/7/20

N2 - West Nile virus (WNV) infections in vertebrates are generally acute but persistent infections have been observed. To investigate the ability of WNV to produce persistent infections, we forced subgenomic WNV replicons to replicate within a cell without causing cell death. Detailed analyses of these cell-adapted genomes revealed mutations within the nonstructural protein genes NS2A (D73H, M108K), NS3 (117Kins), NS4B (E249G) and NS5 (P528H). WNV replicons and WNVs harboring a subset of NS2A or NS3 mutations showed a reduction in genome replication, a reduction in antigen accumulation, a decrease in cytopathic effect, an increased ability to persist in cell culture and/or attenuation in vivo. Taken together, these data indicate that WNV with a defect in replication and an increased potential to persist within the host cell can be generated by point mutations at multiple independent loci, suggesting that persistent viruses could arise in nature.

AB - West Nile virus (WNV) infections in vertebrates are generally acute but persistent infections have been observed. To investigate the ability of WNV to produce persistent infections, we forced subgenomic WNV replicons to replicate within a cell without causing cell death. Detailed analyses of these cell-adapted genomes revealed mutations within the nonstructural protein genes NS2A (D73H, M108K), NS3 (117Kins), NS4B (E249G) and NS5 (P528H). WNV replicons and WNVs harboring a subset of NS2A or NS3 mutations showed a reduction in genome replication, a reduction in antigen accumulation, a decrease in cytopathic effect, an increased ability to persist in cell culture and/or attenuation in vivo. Taken together, these data indicate that WNV with a defect in replication and an increased potential to persist within the host cell can be generated by point mutations at multiple independent loci, suggesting that persistent viruses could arise in nature.

KW - Attenuation

KW - Flavivirus

KW - Innate immune response

KW - Interferon

KW - Nonstructural gene

KW - Persistence

KW - Replication

KW - Replicon

KW - West Nile virus

UR - http://www.scopus.com/inward/record.url?scp=34249722201&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34249722201&partnerID=8YFLogxK

U2 - 10.1016/j.virol.2007.02.009

DO - 10.1016/j.virol.2007.02.009

M3 - Article

C2 - 17382364

AN - SCOPUS:34249722201

SN - 0042-6822

VL - 364

SP - 184

EP - 195

JO - Virology

JF - Virology

IS - 1

ER -

Mutations in West Nile virus nonstructural proteins that facilitate replicon persistence in vitro attenuate virus replication in vitro and in vivo

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this