Home Acta Virologica 2016 Acta Virologica Vol.60, No.3, p.281-289, 2016

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Founded: 1957
ISSN 0001-723X
E-ISSN 1336-2305

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Acta Virologica Vol.60, No.3, p.281-289, 2016

Title: Functional characterization of Bombyx mori nucleopolyhedrovirus mutant lacking late expression factor 9
Author: Y. Zhang, Y. Shi, H. Yu, J. Li, Y. Quan, T. Shu, Z. Nie, Y. Zhang, W. Yu

Abstract: Baculoviridae is a family of invertebrate viruses with large double-stranded DNA genomes. Proteins encoded by some late expression factor (lef ) genes are involved in the regulation of viral gene expression. Lef-9 is one of four transcription-specific Lefs, which are components of the virus-encoded RNA polymerase, and can initiate and transcribe late and very late genes. As a multifunctional protein encoded by the Bombyx mori nucleopolyhedrovirus (BmNPV), Lef-9 may be involved in the regulation of viral propagation. However, the underlying mechanism remains unclear. To determine the role of lef-9 in baculovirus infection, lef-9-knockout virus (lef-9-KO-Bacmid virus) was constructed using the Red recombination system, and the Bac-to-Bac system was used to prepare lef-9-repaired virus (lef-9-Re-Bacmid virus). The lef-9-KO virus did not produce infectious viruses or show infection activity, while the lef-9-repaired virus recovered both. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of the transcription levels in wild-type-Bacmid, lef-9-KO-Bacmid, and lef-9-Re-Bacmid viruses showed that the lef-9-KO bacmid had little effect on viral genome replication. However, the transcription levels of the early and late viral genes, lef-3, ie-1, vp39, and p10, were significantly lower in BmN cells transfected with lef-9-KO-Bacmids than in the controls. Electron microscopy showed no visible enveloped virions in cells transfected with lef-9-KO-Bacmids, while many mature virions in cells transfected with lef-9-Re-Bacmid and wt-Bacmid were present. Thus, lef-9 was not essential for viral genome replication, but significantly affected viral gene transcription and expression in all periods of cell life cycle.

Keywords: Bombyx mori nucleopolyhedrovirus; lef-9 gene; virus replication; virus transcription; virus assembly
Published online: 13-Sep-2016
Year: 2016, Volume: 60, Issue: 3 Page From: 281, Page To: 289

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