title: Glycosylation of the dengue 2 virus E protein at N67 is critical for virus growth in vitro but not for growth in intrathoracically inoculated Aedes aegypti mosquitoes.
abstract: To determine the importance of dengue 2 virus (DEN2V) envelope (E) protein glycosylation, virus mutants in one or both of the N-linked glycosylation motifs were prepared. We found that while the E2 mutant virus (N153Q) replicated in mammalian and mosquito cells, the E1 (N67Q) and E1/2 (N67Q and N153Q) mutant viruses were unable to grow in mammalian cells. Infection of C6/36 mosquito cells with either the E1 or E1/2 mutants resulted in the introduction of a compensatory mutation, K64N, restoring glycosylation in the area. All mutants replicated similarly in inoculated Aedes aegypti mosquitoes, with no change in their mutations. These results suggest that N-linked glycosylation of the E protein is not necessary for DEN2V replication in mosquitoes, however N-linked glycosylation at amino acid N67 (or nearby N64) is critical for the survival of the virus in either mammalian or insect cell culture.
authors.affiliation: Arboviral Diseases Branch, Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services, Fort Collins, CO 80522, USA.
authors.forename: Juliet E
authors.initials: JE
authors.lastname: Bryant
authors.forename: Amanda E
authors.initials: AE
authors.lastname: Calvert
authors.forename: Kyeen
authors.initials: K
authors.lastname: Mesesan
authors.forename: Mary B
authors.initials: MB
authors.lastname: Crabtree
authors.forename: Katharine E
authors.initials: KE
authors.lastname: Volpe
authors.forename: Shawn
authors.initials: S
authors.lastname: Silengo
authors.forename: Richard M
authors.initials: RM
authors.lastname: Kinney
authors.forename: Claire Y-H
authors.initials: CY
authors.lastname: Huang
authors.forename: Barry R
authors.initials: BR
authors.lastname: Miller
authors.forename: John T
authors.initials: JT
authors.lastname: Roehrig
mesh_terms: D000330:Aedes / Q000821:virology*; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002460:Cell Line; D003716:Dengue Virus / Q000235:genetics / Q000254:growth & development*; D006031:Glycosylation; D006801:Humans; D016297:Mutagenesis, Site-Directed; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C051821:E-glycoprotein, Dengue virus type 2; D014759:Viral Envelope Proteins