Characterization of the Interaction Domains between the Phosphoprotein and the Nucleoprotein of Human Metapneumovirus - Université de Versailles Saint-Quentin-en-Yvelines
Article Dans Une Revue Journal of Virology Année : 2022

Characterization of the Interaction Domains between the Phosphoprotein and the Nucleoprotein of Human Metapneumovirus

Résumé

Human metapneumovirus (HMPV) causes severe respiratory diseases in young children. The HMPV RNA genome is encapsidated by the viral nucleoprotein (N), forming an RNA-N complex (NNuc), which serves as the template for genome replication and mRNA transcription by the RNA-dependent RNA polymerase (RdRp). The RdRp is formed by the association of the large polymerase subunit (L), which has RNA polymerase, capping, and methyltransferase activities, and the tetrameric phosphoprotein (P). P plays a central role in the RdRp complex by binding to NNuc and L, allowing the attachment of the L polymerase to the NNuc template. During infection these proteins concentrate in cytoplasmic inclusion bodies (IBs) where viral RNA synthesis occurs. By analogy to the closely related pneumovirus respiratory syncytial virus (RSV), it is likely that the formation of IBs depends on the interaction between HMPV P and NNuc, which has not been demonstrated yet. Here, we finely characterized the binding P-NNuc interaction domains by using recombinant proteins, combined with a functional assay for the polymerase complex activity, and the study of the recruitment of these proteins to IBs by immunofluorescence. We show that the last 6 C-terminal residues of HMPV P are necessary and sufficient for binding to NNuc and that P binds to the N-terminal domain of N (NNTD), and we identified conserved N residues critical for the interaction. Our results allowed us to propose a structural model for the HMPV P-NNuc interaction.
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Dates et versions

hal-03579501 , version 1 (15-10-2021)
hal-03579501 , version 2 (16-11-2022)

Identifiants

Citer

Hortense Decool, Benjamin Bardiaux, Luis Checa Ruano, Olivier Sperandio, Jenna Fix, et al.. Characterization of the Interaction Domains between the Phosphoprotein and the Nucleoprotein of Human Metapneumovirus. Journal of Virology, 2022, 96 (2), pp.e0090921. ⟨10.1128/JVI.00909-21⟩. ⟨hal-03579501v2⟩
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