N-linked glycosylation of the West Nile virus envelope protein is not a requisite for avian virulence or vector competence

Payal D. Maharaj, Stanley A. Langevin, Bethany G. Bolling, Christy C. Andrade, Xavier A. Engle, Wanichaya N. Ramey, Angela Bosco-Lauth, Richard A. Bowen, Todd A. Sanders, Claire Y.H. Huang, William Reisen, Aaron Brault

Research output: Contribution to journalArticle

Abstract

The N-linked glycosylation motif at amino acid position 154–156 of the envelope (E) protein of West Nile virus (WNV) is linked to enhanced murine neuroinvasiveness, avian pathogenicity and vector competence. Naturally occurring isolates with altered E protein glycosylation patterns have been observed in WNV isolates; however, the specific effects of these polymorphisms on avian host pathogenesis and vector competence have not been investigated before. In the present study, amino acid polymorphisms, NYT, NYP, NYF, SYP, SYS, KYS and deletion (A’DEL), were reverse engineered into a parental WNV (NYS) cDNA infectious clone to generate WNV glycosylation mutant viruses. These WNV glycosylation mutant viruses were characterized for in vitro growth, pH-sensitivity, temperature-sensitivity and host competence in American crows (AMCR), house sparrows (HOSP) and Culex quinquefasciatus. The NYS and NYT glycosylated viruses showed higher viral replication, and lower pH and temperature sensitivity than NYP, NYF, SYP, SYS, KYS and A’DEL viruses in vitro. Interestingly, in vivo results demonstrated asymmetric effects in avian and mosquito competence that were independent of the E-protein glycosylation status. In AMCRs and HOSPs, all viruses showed comparable viremias with the exception of NYP and KYS viruses that showed attenuated phenotypes. Only NYP showed reduced vector competence in both Cx. quinquefasciatus and Cx. tarsalis. Glycosylated NYT exhibited similar avian virulence properties as NYS, but resulted in higher mosquito oral infectivity than glycosylated NYS and nonglycosylated, NYP, NYF, SYP and KYS mutants. These data demonstrated that amino acid polymorphisms at E154/156 dictate differential avian host and vector competence phenotypes independent of E-protein glycosylation status.

Original languageEnglish (US)
Article numbere0007473
JournalPLoS neglected tropical diseases
Volume13
Issue number7
DOIs
StatePublished - Jul 1 2019

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Viral Envelope Proteins
West Nile virus
Glycosylation
Mental Competency
Virulence
Viruses
Culicidae
Sparrows
Phenotype
Amino Acids
Crows
Amino Acid Motifs
Culex
Temperature
Viremia
Complementary DNA
Clone Cells
Growth

ASJC Scopus subject areas

  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

Cite this

N-linked glycosylation of the West Nile virus envelope protein is not a requisite for avian virulence or vector competence. / Maharaj, Payal D.; Langevin, Stanley A.; Bolling, Bethany G.; Andrade, Christy C.; Engle, Xavier A.; Ramey, Wanichaya N.; Bosco-Lauth, Angela; Bowen, Richard A.; Sanders, Todd A.; Huang, Claire Y.H.; Reisen, William; Brault, Aaron.

In: PLoS neglected tropical diseases, Vol. 13, No. 7, e0007473, 01.07.2019.

Research output: Contribution to journalArticle

Maharaj, PD, Langevin, SA, Bolling, BG, Andrade, CC, Engle, XA, Ramey, WN, Bosco-Lauth, A, Bowen, RA, Sanders, TA, Huang, CYH, Reisen, W & Brault, A 2019, 'N-linked glycosylation of the West Nile virus envelope protein is not a requisite for avian virulence or vector competence', PLoS neglected tropical diseases, vol. 13, no. 7, e0007473. https://doi.org/10.1371/journal.pntd.0007473
Maharaj, Payal D. ; Langevin, Stanley A. ; Bolling, Bethany G. ; Andrade, Christy C. ; Engle, Xavier A. ; Ramey, Wanichaya N. ; Bosco-Lauth, Angela ; Bowen, Richard A. ; Sanders, Todd A. ; Huang, Claire Y.H. ; Reisen, William ; Brault, Aaron. / N-linked glycosylation of the West Nile virus envelope protein is not a requisite for avian virulence or vector competence. In: PLoS neglected tropical diseases. 2019 ; Vol. 13, No. 7.
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abstract = "The N-linked glycosylation motif at amino acid position 154–156 of the envelope (E) protein of West Nile virus (WNV) is linked to enhanced murine neuroinvasiveness, avian pathogenicity and vector competence. Naturally occurring isolates with altered E protein glycosylation patterns have been observed in WNV isolates; however, the specific effects of these polymorphisms on avian host pathogenesis and vector competence have not been investigated before. In the present study, amino acid polymorphisms, NYT, NYP, NYF, SYP, SYS, KYS and deletion (A’DEL), were reverse engineered into a parental WNV (NYS) cDNA infectious clone to generate WNV glycosylation mutant viruses. These WNV glycosylation mutant viruses were characterized for in vitro growth, pH-sensitivity, temperature-sensitivity and host competence in American crows (AMCR), house sparrows (HOSP) and Culex quinquefasciatus. The NYS and NYT glycosylated viruses showed higher viral replication, and lower pH and temperature sensitivity than NYP, NYF, SYP, SYS, KYS and A’DEL viruses in vitro. Interestingly, in vivo results demonstrated asymmetric effects in avian and mosquito competence that were independent of the E-protein glycosylation status. In AMCRs and HOSPs, all viruses showed comparable viremias with the exception of NYP and KYS viruses that showed attenuated phenotypes. Only NYP showed reduced vector competence in both Cx. quinquefasciatus and Cx. tarsalis. Glycosylated NYT exhibited similar avian virulence properties as NYS, but resulted in higher mosquito oral infectivity than glycosylated NYS and nonglycosylated, NYP, NYF, SYP and KYS mutants. These data demonstrated that amino acid polymorphisms at E154/156 dictate differential avian host and vector competence phenotypes independent of E-protein glycosylation status.",
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AU - Langevin, Stanley A.

AU - Bolling, Bethany G.

AU - Andrade, Christy C.

AU - Engle, Xavier A.

AU - Ramey, Wanichaya N.

AU - Bosco-Lauth, Angela

AU - Bowen, Richard A.

AU - Sanders, Todd A.

AU - Huang, Claire Y.H.

AU - Reisen, William

AU - Brault, Aaron

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