The class II membrane glycoprotein G of bovine respiratory syncytial virus, expressed from a synthetic open reading frame, is incorporated into virions of recombinant bovine herpesvirus 1

Gisela Kuhnle, Astrid Heinze, Jutta Schmitt, Katrin Giesow, Geraldine Taylor, Ivan Morrison, Frans A M Rijsewijk, Jan T Van Oirschot, Gunther M Keil

Research output: Contribution to journalArticlepeer-review

Abstract

The bovine herpesvirus 1 (BHV-1) recombinants BHV-1/eG(ori) and BHV-1/eG(syn) were isolated after insertion of expression cassettes which contained either a genomic RNA-derived cDNA fragment (BHV-1/eG(ori)) or a modified, chemically synthesized open reading frame (ORF) (BHV-1/eG(syn)), which both encode the attachment glycoprotein G of bovine respiratory syncytial virus (BRSV), a class II membrane glycoprotein. Northern blot analyses and nuclear runoff transcription experiments indicated that transcripts encompassing the authentic BRSV G ORF were unstable in the nucleus of BHV-1/eG(ori)-infected cells. In contrast, high levels of BRSV G RNA were detected in BHV-1/eG(syn)-infected cells. Immunoblots showed that the BHV-1/eG(syn)-expressed BRSV G glycoprotein contains N- and O-linked carbohydrates and that it is incorporated into the membrane of infected cells and into the envelope of BHV-1/eG(syn) virions. The latter was also demonstrated by neutralization of BHV-1/eG(syn) infectivity by monoclonal antibodies or polyclonal anti-BRSV G antisera and complement. Our results show that expression of the BRSV G glycoprotein by BHV-1 was dependent on the modification of the BRSV G ORF and indicate that incorporation of class II membrane glycoproteins into BHV-1 virions does not necessarily require BHV-1-specific signals. This raises the possibility of targeting heterologous polypeptides to the viral envelope, which might enable the construction of BHV-1 recombinants with new biological properties and the development of improved BHV-1-based live and inactivated vector vaccines.
Original languageEnglish
Pages (from-to)3804-11
Number of pages8
JournalJ Virol
Volume72
Issue number5
Publication statusPublished - May 1998

Keywords

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cattle
  • Cell Line
  • DNA, Viral
  • Gene Expression
  • Genetic Vectors/genetics
  • Genome, Viral
  • Herpesvirus 1, Bovine/genetics
  • Molecular Sequence Data
  • Neutralization Tests
  • Open Reading Frames
  • Recombinant Fusion Proteins
  • Recombination, Genetic
  • Respiratory Syncytial Virus, Bovine/genetics
  • Respiratory Syncytial Virus, Bovine/metabolism
  • Transcription, Genetic
  • Viral Envelope Proteins/biosynthesis
  • Viral Envelope Proteins/genetics
  • Virion/metabolism

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