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

PLoS One. 2015 Dec 14;10(12):e0144930. doi: 10.1371/journal.pone.0144930. eCollection 2015.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bombyx / virology
  • Cell Line
  • DNA, Viral / metabolism
  • Gene Knockout Techniques
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Nucleopolyhedroviruses / genetics
  • Nucleopolyhedroviruses / physiology*
  • Protein Multimerization
  • Real-Time Polymerase Chain Reaction
  • Viral Proteins / chemistry
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*
  • Virus Replication

Substances

  • DNA, Viral
  • Viral Proteins

Grants and funding

This work was supported by grants from the National High-Tech R&D Program of China (No. 2013AA102507), the National Natural Science Foundation of China (Nos. 31172269, 31272505 and 31472152), the China Agriculture Research System (CARS-22) and Fundamental Research Funds for the Central Universities (XDJK2015D002).