Structural Dynamics of the C-terminal X Domain of Nipah and Hendra Viruses Controls the Attachment to the C-terminal Tail of the Nucleocapsid Protein. Issue 10 (30th May 2022)

• Record Type: Journal Article
• Authors: Bourhis, Jean-Marie; Yabukarski, Filip; Communie, Guillaume; Schneider, Robert; Volchkova, Valentina A.; Frénéat, Mickaël; Gérard, Francine C.; Ducournau, Corinne; Mas, Caroline; Tarbouriech, Nicolas; Ringkjøbing Jensen, Malene; Volchkov, Viktor E.; Blackledge, Martin; Jamin, Marc
• Is Part Of: Journal of molecular biology. Volume 434:Issue 10(2022)
• Publication Date: 2022-05-30
• Subjects: nipah virus -- phosphoprotein -- mononegavirales -- intrinsically disordered protein -- small-angle X-ray scattering; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2022.167551
• Languages: English
• ISSNs: 0022-2836

Abstract

Graphical abstract: Highlights: X domain of henipavirus P protein (PXD ) is a key element in the replication complex. Nipah virus PXD exchanges between different conformations and folds upon binding N. Nipah virus PXD but not Hendra virus PXD forms dimers. Recombinant NiV virus with PXD exchanged for HeV PXD was slightly attenuated. Tuning of PXD stability is a way to regulate the attachment of P to the nucleocapsid. Abstract: To understand the dynamic interactions between the phosphoprotein (P) and the nucleoprotein (N) within the transcription/replication complex of the Paramyxoviridae and to decipher their roles in regulating viral multiplication, we characterized the structural properties of the C-terminal X domain (PXD ) of Nipah (NiV) and Hendra virus (HeV) P protein. In crystals, isolated NiV PXD adopted a two-helix dimeric conformation, which was incompetent for binding its partners, but in complex with the C-terminal intrinsically disordered tail of the N protein (NTAIL ), it folded into a canonical 3H bundle conformation. In solution, SEC-MALLS, SAXS and NMR spectroscopy experiments indicated that both NiV and HeV PXD were larger in size than expected for compact proteins of the same molecular mass and were in conformational exchange between a compact three-helix (3H) bundle and partially unfolded conformations, where helix α3 is detached from the other two. Some measurements also provided strong evidence for dimerization of NiV PXD in solution but not for HeV PXD . Ensemble modeling of experimental SAXS data and statistical-dynamical modeling reconciled all these data, yielding a model where NiV and HeV PXD exchanged between different conformations, and where NiV but not HeV PXD formed dimers. Finally, recombinant NiV comprising a chimeric P carrying HeV PXD was rescued and compared with parental NiV. Experiments carried out in cellula demonstrated that the replacement of PXD did not significantly affect the replication dynamics while caused a slight virus attenuation, suggesting a possible role of the dimerization of NiV PXD in viral replication.