title: The baculovirus core gene ac83 is required for nucleocapsid assembly and per os infectivity of Autographa californica nucleopolyhedrovirus.
abstract: Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac83 is a baculovirus core gene whose function in the AcMNPV life cycle is unknown. In the present study, an ac83-knockout AcMNPV (vAc83KO) was constructed to investigate the function of ac83 through homologous recombination in Escherichia coli. No budded virions were produced in vAc83KO-transfected Sf9 cells, although viral DNA replication was unaffected. Electron microscopy revealed that nucleocapsid assembly was aborted due to the ac83 deletion. Domain-mapping studies revealed that the expression of Ac83 amino acid residues 451 to 600 partially rescued the ability of AcMNPV to produce infectious budded virions. Bioassays indicated that deletion of the chitin-binding domain of Ac83 resulted in the failure of oral infection of Trichoplusia ni larvae by AcMNPV, but AcMNPV remained infectious following intrahemocoelic injection, suggesting that the domain is involved in the binding of occlusion-derived virions to the peritrophic membrane and/or to other chitin-containing insect tissues. It has been demonstrated that Ac83 is the only component with a chitin-binding domain in the per os infectivity factor complex on the occlusion-derived virion envelope. Interestingly, a functional inner nuclear membrane sorting motif, which may facilitate the localization of Ac83 to the envelopes of occlusion-derived virions, was identified by immunofluorescence analysis. Taken together, these results demonstrate that Ac83 plays an important role in nucleocapsid assembly and the establishment of oral infection.
authors.affiliation: State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China.
authors.forename: Shimao
authors.initials: S
authors.lastname: Zhu
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Meijin
authors.initials: M
authors.lastname: Yuan
authors.forename: Kai
authors.initials: K
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000502:physiology*; D002478:Cells, Cultured; D002686:Chitin / Q000378:metabolism; D019295:Computational Biology; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000378:metabolism*; D017924:Nucleopolyhedroviruses / Q000472:pathogenicity*; D017434:Protein Structure, Tertiary; D060888:Real-Time Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D014758:Viral Core Proteins / Q000235:genetics* / Q000378:metabolism; D019065:Virus Assembly; D014779:Virus Replication
chemical_list: D004279:DNA, Viral; D014758:Viral Core Proteins; D002686:Chitin