title: Baculovirus VP80 protein and the F-actin cytoskeleton interact and connect the viral replication factory with the nuclear periphery.
abstract: Recently, we showed that the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) VP80 protein is essential for the formation of both virion types, budded virus (BV) and occlusion-derived virus (ODV). Deletion of the vp80 gene did not affect assembly of nucleocapsids. However, these nucleocapsids were not able to migrate from the virogenic stroma to the nuclear periphery. In the current paper, we constructed a baculovirus recombinant with enhanced-green fluorescent protein (EGFP)-tagged VP80, allowing visualization of the VP80 distribution pattern during infection. In baculovirus-infected cells, the EGFP-VP80 protein is entirely localized in nuclei, adjacent to the virus-triggered F-actin scaffold that forms a highly organized three-dimensional network connecting the virogenic stroma physically with the nuclear envelope. Interaction between VP80 and host actin was confirmed by coimmunoprecipitation. We further showed that VP80 is associated with the nucleocapsid fraction of both BVs and ODVs, typically at one end of the nucleocapsids. In addition, the presence of sequence motifs with homology to invertebrate paramyosin proteins strongly supports a role for VP80 in the polar transport of nucleocapsids to the periphery of the nucleus on their way to the plasma membrane to form BVs and for assembly in the nuclear periphery to form ODVs for embedding in viral occlusion bodies.
authors: Laboratory of Virology, Wageningen University, PB Wageningen, The Netherlands.
authors: Martin
authors: M
authors: Marek
authors: Otto-Wilhelm
authors: OW
authors: Merten
authors: Lionel
authors: L
authors: Galibert
authors: Just M
authors: JM
authors: Vlak
authors: Monique M
authors: MM
authors: van Oers
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D019677:Artificial Gene Fusion; D036022:Capsid Proteins / Q000378:metabolism*; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D054884:Host-Pathogen Interactions; D047468:Immunoprecipitation; D007915:Lepidoptera; D017924:Nucleopolyhedroviruses / Q000472:pathogenicity*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000199:Actins; D036022:Capsid Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C076630:Ac-vp80 protein, Autographa californica nucleopolyhedrovirus