Both E protein glycans adversely affect dengue virus infectivity but are beneficial for virion release.
The E protein of most flaviviruses is modified by Asn-linked glycosylation at residue 153/154 and in the case of the four dengue virus (DENV) serotypes by a second glycan at residue 67. However, the absence of E protein glycosylation among numerous natural isolates of different flaviviruses suggests that the glycan, per se, is not critically important in the virus life cycle. Consistent with this notion, we show that ablation of both glycans from the DENV-2 E protein reduces but does not prevent growth of the variant in mammalian and mosquito cells. We found a pronounced and opposing effect of glycan ablation on two stages of the virus growth cycle: infectivity and release. Loss of either of the two DENV E protein glycans markedly enhanced infectivity of variants for mosquito cells at the expense of efficient virion release. The variants also displayed reduced release in mammalian cells, which was more prominent for viruses lacking the Asn 67-linked glycan than for those lacking the Asn 153-linked glycan, without a marked change in infectivity. Mutations, which compensated for the defect in virus morphogenesis associated with ablation of the Asn 67-linked glycan in mammalian cells but interestingly not in mosquito cells, were identified at the glycosylation acceptor motif and a second site in E protein domain II. The dueling influences of infectivity and release on virus growth affected by the glycans may explain the plasticity in E protein glycosylation among the flaviviruses.
Lee
Eva
E
John Curtin School of Medical Research, the Australian National University, Canberra, ACT, Australia.
Leang
Sook Kwan
SK
Davidson
Andrew
A
Lobigs
Mario
M
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D009033:Culicidae; D003716:Dengue Virus / Q000472:pathogenicity* / Q000502:physiology*; D020125:Mutation, Missense; D013489:Suppression, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics / Q000502:physiology*; D014774:Virulence; D037521:Virulence Factors / Q000502:physiology*; D057074:Virus Release
C051821:E-glycoprotein, Dengue virus type 2; D014759:Viral Envelope Proteins; D037521:Virulence Factors