A baculovirus single-stranded DNA binding protein, LEF-3, mediates the nuclear localization of the putative helicase P143.
The intracellular localization of the baculovirus Autographa californica nuclear polyhedrosis virus p143 gene product (P143) was investigated by immunofluoresence staining of infected and transfected cells. As expected for a protein essential for viral DNA replication, under these conditions, P143 was localized to the nucleus. However, when a plasmid directing the synthesis of P143 from its own promoter was co-transfected with a plasmid expressing IE-1, P143 was found in the cytoplasm. Cotransfection of cells with a series of plasmids expressing viral genes sufficient for stimulation of plasmid replication resulted in nuclear localization of P143 suggesting that one of the gene products required for viral DNA replication may also assist nuclear localization of P143. Sequential deletion of various genes from this assay revealed that when the plasmid expressing LEF-3 was deleted from the mixture, P143 remained in the cytoplasm. Plasmids were constructed where the synthesis of LEF-3 and P143 were directed by the ie-1 promoter. When these plasmids were cotransfected, both LEF-3 and P143 colocalized to the nucleus suggesting that an important function of LEF-3 is intracellular trafficking of P143 and directing it to the nucleus.
Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada.
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Wu
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Carstens
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D018411:Spodoptera; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D017931:DNA Primers; D004268:DNA-Binding Proteins; D014764:Viral Proteins; C094647:late expression factor 3, baculovirus; D004265:DNA Helicases