title: Essential C-terminal region of the baculovirus minor capsid protein VP80 binds DNA.
abstract: The essential Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) minor capsid protein VP80 has been recently shown to interact with the virus-triggered, nuclear F-actin cytoskeleton. A role for VP80 in virus morphogenesis has been proposed in the maturation of progeny nucleocapsids and in their egress from the virogenic stroma toward the nuclear periphery by a mechanism, which also includes F-actin filaments. We performed functional mapping of VP80 demonstrating that its highly conserved C-terminal region plays a crucial role in virion morphogenesis. Protein database mining identified a putative basic helix-loop-helix (bHLH) domain, a DNA-binding module typical for eukaryotic transcription factors, in the essential C-terminal region of VP80. Using a molecular modeling approach, we predicted the three-dimensional structure of this domain, revealing some unique properties. Biochemical assays proved that VP80 can form homodimers, a critical prerequisite of DNA-binding bHLH proteins. The ability of VP80 to bind DNA was subsequently confirmed by an electrophoretic mobility shift assay. We further show that AcMNPV DNA replication occurs in the absence of VP80. Immunolabeling of VP80 in baculovirus-infected cells rather points toward its involvement in nucleocapsid maturation. The competence of VP80 to interact with both F-actin and DNA provides novel insight into baculovirus morphogenesis.
authors.affiliation: Laboratory of Virology, Wageningen University, Wageningen, Netherlands.
authors.forename: Martin
authors.initials: M
authors.lastname: Marek
authors.forename: Otto-Wilhelm
authors.initials: OW
authors.lastname: Merten
authors.forename: Feana
authors.initials: F
authors.lastname: Francis-Devaraj
authors.forename: Monique M van
authors.initials: MM
authors.lastname: Oers
mesh_terms: D000595:Amino Acid Sequence; D016367:Baculoviridae / Q000737:chemistry*; D001483:Base Sequence; D036022:Capsid Proteins / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D017931:DNA Primers; D024202:Electrophoretic Mobility Shift Assay; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
chemical_list: D036022:Capsid Proteins; D017931:DNA Primers; D004247:DNA
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