Isolation and characterisation of the rabies virus N°-P complex produced in insect cells

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Manos Mavrakis - , European Molecular Biology Laboratory (EMBL) Heidelberg (Author)
  • Frédéric Iseni - , European Molecular Biology Laboratory (EMBL) Heidelberg, University of Geneva (Author)
  • Catherine Mazza - , European Molecular Biology Laboratory (EMBL) Heidelberg (Author)
  • Guy Schoehn - , European Molecular Biology Laboratory (EMBL) Heidelberg, Université Grenoble Alpes (Author)
  • Christine Ebel - , Université Grenoble Alpes (Author)
  • Marc Gentzel - , European Molecular Biology Laboratory (EMBL) Heidelberg (Author)
  • Thomas Franz - , European Molecular Biology Laboratory (EMBL) Heidelberg (Author)
  • Rob W.H. Ruigrok - , European Molecular Biology Laboratory (EMBL) Heidelberg, Université Grenoble Alpes (Author)

Abstract

When the nucleoprotein (N) of nonsegmented negative-strand RNA viruses is expressed in insect cells, it binds to cellular RNA and forms N-RNA complexes just like viral nucleocapsids. However, in virus-infected cells, N is prevented from binding to cellular RNA because a soluble complex is formed between N and the viral phosphoprotein (P), the N°-P complex. N is only released from this complex for binding to newly made viral or complementary RNA. We coexpressed rabies virus N and P proteins in insect cells and purified the N°-P complex. Characterisation by gel filtration, polyacrylamide gel electrophoresis, analytical ultracentrifugation, native mass spectroscopy, and electron microscopy showed that the complex consists of one N protein plus two P proteins, i.e., an N°-P2 complex.

Details

Original languageEnglish
Pages (from-to)406-414
Number of pages9
JournalVirology
Volume305
Issue number2
Publication statusPublished - 20 Jan 2003
Peer-reviewedYes
Externally publishedYes

External IDs

PubMed 12573586
ORCID /0000-0002-4482-6010/work/142251047

Keywords

ASJC Scopus subject areas