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Scientific article
Open access
English

A Sendai virus vector leading to the efficient expression of mutant M proteins interfering with virus particle budding

Published inVirology, vol. 221, no. 1, p. 159-171
Publication date1996
Abstract

A Sendai virus expression vector in the form of a transcribing copy-back defective interfering RNA was constructed and shown to efficiently express a tagged matrix protein in the only context of a Sendai virus infection. In an attempt to identify relevant M protein domains involved in viral assembly and budding, a series of deletion mutants were tested for their ability to bind to cellular membrane fractions. The deletion of a region spanning amino acids 105-137 significantly decreased this binding when the protein was expressed in a system driven by the T7 RNA polymerase away from any other viral proteins. Plus or minus charges were introduced in the hydrophobic portion of a predicted amphiphilic helix in this region, and M proteins with altered membrane binding properties were produced. The genes encoding these mutant M proteins were then inserted in the Sendai virus vector and shown to be expressed at levels similar to that of the endogenous wild-type M protein. The presence of a negative charge in the hydrophobic region of the putative amphiphilic helix prevented the incorporation of the mutant protein into virus particles and appeared to decrease the efficiency of virus particle budding. In contrast, the introduction of a positive charge appeared to increase the M mutant uptake into virions. The use a Sendai virus vector has therefore been shown instrumental in the identification of mutant M proteins interfering with the viral assembly-budding process.

Keywords
  • Animals
  • Base Sequence
  • Cell Line
  • Cell Membrane/metabolism
  • Cloning
  • Molecular
  • Conserved Sequence
  • Cricetinae
  • DNA
  • Viral
  • Genetic Vectors/genetics
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Parainfluenza Virus 1
  • Human/genetics/physiology
  • Point Mutation
  • Rabbits
  • Sequence Deletion
  • Viral Matrix Proteins/genetics/physiology
  • Virus Assembly/physiology
Citation (ISO format)
MOTTET-OSMAN, Geneviève et al. A Sendai virus vector leading to the efficient expression of mutant M proteins interfering with virus particle budding. In: Virology, 1996, vol. 221, n° 1, p. 159–171. doi: 10.1006/viro.1996.0362
Main files (1)
Article (Published version)
Identifiers
ISSN of the journal0042-6822
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