Characterization of Spodoptera litura multicapsid nucleopolyhedrovirus 38.7 k protein, which contains a conserved BRO domain

Virus Res. 2006 Feb;115(2):185-91. doi: 10.1016/j.virusres.2005.08.006. Epub 2005 Oct 3.

Abstract

Homology analysis revealed that Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) 38.7 k protein has 22-83% amino acid identities with Ecotropis obliqua NPV, Mamestra configurata MNPV, Helicoverpa armigera SNPV, H. zea SNPV, S. exigua MNPV and S. littoralis MNPV 38.7 k proteins. Analysis of the relationship of these 38.7 k proteins indicated that they contain a conserved BRO-N domain, and SpltMNPV and SpliMNPV 38.7 k proteins also contain a motif found in all known viral and prokaryotic single-strand DNA binding proteins. RT-PCR results showed that SpltMNPV 38.7 k gene is transcribed actively at the late stage of infection and the mRNA start site was mapped within a consensus baculovirus late promoter motif (ATAAG). Western blot analysis revealed that the 38.7 k was expressed in infected S. litura cells as a 41 kDa form and this protein distributed in the nucleus of infected cells. Using a histone extraction protocol, SpltMNPV 38.7 k could be detected in the histone H1 fraction. Micrococcal nuclease treatment released SpltMNPV 38.7 k protein from the chromatin fraction, suggesting that its involvement in nucleosome structures. Furthermore, column chromatography using DNA-cellulose showed that SpltMNPV 38.7 k protein interacted with nucleic acids. It was proposed that SpltMNPV 38.7 k might function as a DNA-binding protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs / genetics
  • Animals
  • Blotting, Western
  • Cell Line
  • Cell Nucleus / chemistry
  • DNA / metabolism
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / isolation & purification
  • Gene Expression
  • Histones / isolation & purification
  • Nucleopolyhedroviruses / chemistry
  • Nucleopolyhedroviruses / genetics
  • Nucleopolyhedroviruses / physiology*
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA, Messenger / analysis
  • RNA, Viral
  • Sequence Homology, Amino Acid
  • Spodoptera / virology*
  • Transcription Initiation Site
  • Viral Proteins / chemistry*
  • Viral Proteins / genetics*
  • Viral Proteins / isolation & purification
  • Viral Proteins / physiology

Substances

  • DNA-Binding Proteins
  • Histones
  • RNA, Messenger
  • RNA, Viral
  • Viral Proteins
  • DNA