title: Role of N-glycosylation on Zika virus E protein secretion, viral assembly and infectivity.
abstract: Zika virus has rapidly spread reaching a global distribution pattern similar to that of dengue virus, and has been associated with serious neurological and developmental pathologies, like congenital malformation during pregnancy and Guillain-Barré syndrome. Sequence analysis of different clinical and laboratory isolates has shown the existence of mutants with loss of the conserved N-glycosylation motif on domain I of protein E that is common to all flaviviruses. We found that loss of E N-linked glycosylation leads to compromised expression and secretion of E ectodomain from mammalian cells. For both, wild type and glycosylation-negative mutant, secretion was independent of co-expression of the PrM viral protein, but highly dependent on temperature. Low temperature (28 °C) favoured secretion, although the glycosylation mutant E ectodomain showed impaired secretion and membrane display compared to the wild type. Production of pseudoviral particles with a West Nile virus replicon packaged with the Zika virus structural proteins C-PrM-E was significantly reduced with the non-glycosylated E. Similarly, glycosylation-negative pseudoviral particles showed impaired infectivity of Vero cells and reduced ability to infect K562 cells upon particles opsonisation with anti-E antibodies.
authors.affiliation: Molecular Immunology Group, International Centre for Genetic Engineering and Biotechnology, 34149 Trieste, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Mossenta
authors.affiliation: Molecular Immunology Group, International Centre for Genetic Engineering and Biotechnology, 34149 Trieste, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Marchese
authors.affiliation: Molecular Immunology Group, International Centre for Genetic Engineering and Biotechnology, 34149 Trieste, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Poggianella
authors.affiliation: Molecular Immunology Group, International Centre for Genetic Engineering and Biotechnology, 34149 Trieste, Italy. Electronic address: slon@icgeb.org.
authors.forename: J L
authors.initials: JL
authors.lastname: Slon Campos
authors.affiliation: Molecular Immunology Group, International Centre for Genetic Engineering and Biotechnology, 34149 Trieste, Italy. Electronic address: burrone@icgeb.org.
authors.forename: O R
authors.initials: OR
authors.lastname: Burrone
mesh_terms: D000818:Animals; D006031:Glycosylation; D006801:Humans; D020014:K562 Cells; D000072417:Protein Domains; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014775:Virus Activation / Q000502:physiology*; D019065:Virus Assembly / Q000502:physiology*; D000071244:Zika Virus / Q000472:pathogenicity / Q000502:physiology*; D000071243:Zika Virus Infection / Q000821:virology*
chemical_list: D014759:Viral Envelope Proteins
keywords: Envelope protein; Flavivirus; Glycosylation; Infectivity; Secretion; Zika virus