Characterization of the interaction domains between the phosphoprotein and the nucleocapsid of human Metapneumovirus - Archive ouverte HAL Access content directly
Preprints, Working Papers, ... Year :

Characterization of the interaction domains between the phosphoprotein and the nucleocapsid of human Metapneumovirus

(1) , (2) , (2) , (2, 3) , (1) , (4) , (1) , (1) , (1) , (1)
1
2
3
4

Abstract

Human metapneumovirus (HMPV) causes severe respiratory diseases in young children. The HMPV RNA genome is encapsidated by the viral nucleoprotein, forming an RNA-N complex (N Nuc ), which serves as 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 N Nuc and L, allowing the attachment of the L polymerase to the nucleocapsid 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 P and N Nuc . However, the HMPV P-N Nuc interaction still remains to characterize. Here, we finely characterized the binding domains involved in HMPV P and N Nuc interaction by studying binding between recombinant proteins, combined with the use of a functional assay of 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 N Nuc , that P binds the N-terminal domain of N (N NTD ), and identified conserved N residues critical for the interaction. Our results allowed to propose a structural model of the HMPV P-N Nuc interaction. IMPORTANCE Like respiratory syncytial virus (RSV), human metapneumovirus (HMPV) is a leading cause of severe respiratory infections in children but also affects human populations of all ages worldwide. Nowadays, no vaccine or efficient antiviral treatments are available for these two pneumoviruses. A better understanding of the molecular mechanisms involved in viral replication could help the design or discovery of specific antiviral compounds. In this work we have investigated the interaction between two major viral proteins involved in HMPV RNA synthesis, the N and P proteins. We finely characterized their domains of interaction, in particular a pocket on the surface of the N protein that could be used as a template for the design of compounds interfering with N-P complexes and viral replication.
Fichier principal
Vignette du fichier
2021.06.02.446795v1.full.pdf (1.49 Mo) Télécharger le fichier
Origin : Files produced by the author(s)

Dates and versions

pasteur-03380015 , version 1 (15-10-2021)

Licence

Attribution - NonCommercial - NoDerivatives - CC BY 4.0

Identifiers

Cite

Hortense Decool, Benjamin Bardiaux, Luis Checa Ruano, Olivier Sperandio, Jenna Fix, et al.. Characterization of the interaction domains between the phosphoprotein and the nucleocapsid of human Metapneumovirus. 2021. ⟨pasteur-03380015⟩
76 View
39 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More