title: Characterization of protein-protein interaction domains within the baculovirus Autographa californica multiple nucleopolyhedrovirus late expression factor LEF-3.
abstract: Autographa californica nucleopolyhedrovirus late expression factor 3 (LEF-3) is required for late viral gene expression probably through its numerous functions related to DNA replication, including nuclear localization of the virus helicase P143 and binding to ssDNA. LEF-3 appears to interact with itself as a homo-oligomer, although the details of this oligomeric structure are not yet known. To examine LEF-3-LEF-3 interactions, a bimolecular fluorescent protein complementation assay was used. Pairs of recombinant plasmids expressing full-length LEF-3 fused to one of two complementary fragments (V1 or V2) of a variant of yellow fluorescent protein named 'Venus' were constructed. Plasmids expressing fusions with complementary fragments of Venus were co-transfected into Sf21 cells and analysed by fluorescence microscopy. Co-transfected plasmids expressing full-length V1-LEF-3 and V2-LEF-3 showed positive fluorescence, confirming the formation of homo-oligomers. A series of truncated V1/V2-LEF-3 fusions was constructed and used to investigate interactions with one another as well as with full-length LEF-3.
authors.affiliation: Department of Biomedical and Molecular Sciences Queen's University Kingston, ON K7L 3N6 Canada.
authors.forename: Kelsey
authors.initials: K
authors.lastname: Downie
authors.affiliation: Department of Biomedical and Molecular Sciences Queen's University Kingston, ON K7L 3N6 Canada.
authors.forename: Gbolagade
authors.initials: G
authors.lastname: Adetola
authors.affiliation: Department of Biomedical and Molecular Sciences Queen's University Kingston, ON K7L 3N6 Canada.
authors.forename: Eric B
authors.initials: EB
authors.lastname: Carstens
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007915:Lepidoptera / Q000821:virology*; D008856:Microscopy, Fluorescence; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D054730:Protein Interaction Domains and Motifs / Q000235:genetics*; D018411:Spodoptera / Q000821:virology; D014162:Transfection; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D014764:Viral Proteins; C094647:late expression factor 3, baculovirus