Oligomerization of Baculovirus LEF-11 Is Involved in Viral DNA Replication

We have previously reported that baculovirus Bombyx mori nucleopolyhedrovirus (BmNPV) late expression factor 11 (lef-11) is associated with viral DNA replication and have demonstrated that it potentially interacts with itself; however, whether LEF-11 forms oligomers and the impact of LEF-11 oligomer...

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Bibliographic Details
Published in:PloS one 2015-12, Vol.10 (12), p.e0144930
Main Authors: Dong, Zhan-Qi, Hu, Nan, Zhang, Jun, Chen, Ting-Ting, Cao, Ming-Ya, Li, Hai-Qing, Lei, Xue-Jiao, Chen, Peng, Lu, Cheng, Pan, Min-Hui
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analysis
Animals
Baculovirus
Bombyx - virology
Bombyx mori
Cell Line
Deoxyribonucleic acid
DNA
DNA biosynthesis
DNA replication
DNA, Viral - metabolism
Gene expression
Gene Knockout Techniques
Genetic Vectors - genetics
Genetic Vectors - metabolism
Genomics
Hydrophobicity
Laboratories
Microscopy, Fluorescence
Molecular Sequence Data
Mutagenesis, Site-Directed
Mutants
Nucleopolyhedrovirus - genetics
Nucleopolyhedrovirus - physiology
Oligomerization
Oligomers
Plasmids
Protein Multimerization
Real-Time Polymerase Chain Reaction
Replication
Site-directed mutagenesis
Viral Proteins - chemistry
Viral Proteins - genetics
Viral Proteins - metabolism
Virus Replication
Viruses
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Summary:We have previously reported that baculovirus Bombyx mori nucleopolyhedrovirus (BmNPV) late expression factor 11 (lef-11) is associated with viral DNA replication and have demonstrated that it potentially interacts with itself; however, whether LEF-11 forms oligomers and the impact of LEF-11 oligomerization on viral function have not been substantiated. In this study, we first demonstrated that LEF-11 is capable of forming oligomers. Additionally, a series of analyses using BmNPV LEF-11 truncation mutants indicated that two distinct domains control LEF-11 oligomerization (aa 42-61 and aa 72-101). LEF-11 truncation constructs were inserted into a lef-11-knockout BmNPV bacmid, which was used to demonstrate that truncated LEF-11 lacking either oligomerization domain abrogates viral DNA replication. Finally, site-directed mutagenesis was used to determine that the conserved hydrophobic residues Y58&I59 (representing Y58 and I59), I85 and L88&L89 (representing L88 and L89) are required for LEF-11 oligomerization and viral DNA replication. Collectively, these data indicate that BmNPV LEF-11 oligomerization influences viral DNA replication.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0144930