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Media type:
E-Article
Title:
Autographa californica Multiple Nucleopolyhedrovirus LEF-2 Is a Capsid Protein Required for Amplification but Not Initiation of Viral DNA Replication
Description:
<jats:title>ABSTRACT</jats:title>
<jats:p>
The late expression factor 2 gene (
<jats:italic>lef-2</jats:italic>
) of baculovirus
<jats:italic>Autographa californica</jats:italic>
multiple nucleopolyhedrovirus (AcMNPV) has been identified as one of the factors essential for origin-dependent DNA replication in transient expression assays and has been shown to be involved in late/very late gene expression. To study the function of
<jats:italic>lef-2</jats:italic>
in the life cycle of AcMNPV,
<jats:italic>lef-2</jats:italic>
knockout and repair bacmids were generated by homologous recombination in
<jats:italic>Escherichia coli</jats:italic>
. Growth curve analysis showed that
<jats:italic>lef-2</jats:italic>
was essential for virus production. Interestingly, a DNA replication assay indicated that
<jats:italic>lef-2</jats:italic>
is not required for the initiation of viral DNA replication and that, rather, it is required for the amplification of DNA replication.
<jats:italic>lef-2</jats:italic>
is also required for the expression of late and very late genes, as the expression of these genes was abolished by
<jats:italic>lef-2</jats:italic>
deletion. Temporal and spatial distributions of LEF-2 protein in infected cells were also analyzed, and the data showed that LEF-2 protein was localized to the virogenic stroma in the nuclei of the infected cells. Analysis of purified virus particles revealed that LEF-2 is a viral protein component of both budded and occlusion-derived virions, predominantly in the nucleocapsids of the virus particles. This observation suggests that LEF-2 may be required immediately after virus entry into host cells for efficient viral DNA replication.
</jats:p>