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

Article Article scientifique Article A Sendai virus vector leading to the efficient expression of mutant M proteins interfering with virus particle budding eng 1996 author Mottet-Osman Geneviève 26297 author Muehlemann Andréas Samuel 115387 author Tapparel Caroline 0000-0002-0411-6567 27039 author Hoffmann Frédéric Olivier 23361 author Roux Laurent 24751 159-171 10.1006/viro.1996.0362 8661424 0042-6822 https://www.sciencedirect.com/science/article/pii/S0042682296903621 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. 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 Dewey 616 54 Département de microbiologie et médecine moléculaire 21B false 931 Identification des facteurs clés impliqués dans la pathogénèse des rhinovirus et entérovirus 953 Virology 221 1 934263c6-b097-415f-962a-344323393990 Article (Published version) Public 1-s2.0-S0042682296903621-main.pdf 354521 application/pdf OA OTHER openAccess Faculté de médecine / Section de médecine fondamentale / Département de microbiologie et médecine moléculaire

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A Sendai virus vector leading to the efficient expression of mutant M proteins interfering with virus particle budding	Virology			1996	436	590

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Hoffmann, Frédéric Olivier