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Dimerization efficiency of canine distemper virus matrix protein regulates membrane-budding activity

Type of publication Peer-reviewed
Publikationsform Original article (peer-reviewed)
Author Bringolf Fanny, Bringolf Fanny, Herren Michael, Herren Michael, Wyss Marianne, Vidondo Beatriz, Langedijk Johannes P., Zurbriggen Andreas, Plattet Philippe,
Project Paramyxovirus cell exit: from basics to drug discovery strategies
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Original article (peer-reviewed)

Journal Journal of Virology
Volume (Issue) 91(16)
Page(s) e00521-17 - e00521-17
Title of proceedings Journal of Virology
DOI 10.1128/jvi.00521-17

Abstract

© 2017 American Society for Microbiology. Paramyxoviruses rely on the matrix (M) protein to orchestrate viral assembly and budding at the plasma membrane. Although the mechanistic details remain largely unknown, structural data suggested that M dimers and/or higher-order oligomers may facilitate membrane budding. To gain functional insights, we employed a structure-guided mutagenesis approach to investigate the role of canine distemper virus (CDV) M protein self-assembly in membrane-budding activity. Three six-alanine-block (6A-block) mutants with mutations located at strategic oligomeric positions were initially designed. While the first one includes residues potentially residing at the protomer-protomer interface, the other two display amino acids located within two distal surface-exposed α-helices proposed to be involved in dimerdimer contacts. We further focused on the core of the dimeric interface by mutating asparagine 138 (N138) to several nonconservative amino acids. Cellular localization combined with dimerization and coimmunopurification assays, performed under various denaturing conditions, revealed that all 6A-block mutants were impaired in selfassembly and cell periphery accumulation. These phenotypes correlated with deficiencies in relocating CDV nucleocapsid proteins to the cell periphery and in virus-like particle (VLP) production. Conversely, all M-N138 mutants remained capable of selfassembly, though to various extents, which correlated with proper accumulation and redistribution of nucleocapsid proteins at the plasma membrane. However, membrane deformation and VLP assays indicated that the M-N138 variants exhibiting the most reduced dimerization propensity were also defective in triggering membrane remodeling and budding, despite proper plasma membrane accumulation. Overall, our data provide mechanistic evidence that the efficiency of CDV M dimerization/oligomerization governs both cell periphery localization and membrane-budding activity.
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