Atomic Resolution Description of the Interaction between the Nucleoprotein and Phosphoprotein of Hendra Virus

PLoS Pathogens

Johnny Habchi¹  Nicolas Papageorgiou²  Sonia Longhi²  Filip Yabukarski³  Robert Schneider⁴  Marc Jamin⁴  Nicolas Tarbouriech⁵  Malene Ringkjøbing Jensen⁵  Martin Blackledge⁶  Guillaume Communie⁶  Rob W. H. Ruigrok⁷  David Blocquel⁷ 

[1] CEA, DSV, IBS, Grenoble, France;CNRS and Aix-Marseille Université, Architecture et Fonction des Macromolécules Biologiques, UMR 7257, Marseille, France;CNRS, IBS, Grenoble, France;CNRS, UVHCI, Grenoble, France;Unit for Virus Host Cell Interactions, Université Grenoble Alpes-EMBL-CNRS, Grenoble, France;Université Grenoble Alpes, Institut de Biologie Structurale (IBS), Grenoble, France;Université Grenoble Alpes, UVHCI, Grenoble, France

关键词: Nucleocapsids;    Crystal structure;    NMR spectroscopy;    Polymerases;    Paramyxoviruses;    Protein structure;    Intrinsically disordered proteins;    Nucleoproteins;   

DOI  :  10.1371/journal.ppat.1003631

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Hendra virus (HeV) is a recently emerged severe human pathogen that belongs to the Henipavirus genus within the Paramyxoviridae family. The HeV genome is encapsidated by the nucleoprotein (N) within a helical nucleocapsid. Recruitment of the viral polymerase onto the nucleocapsid template relies on the interaction between the C-terminal domain, NTAIL, of N and the C-terminal X domain, XD, of the polymerase co-factor phosphoprotein (P). Here, we provide an atomic resolution description of the intrinsically disordered NTAIL domain in its isolated state and in intact nucleocapsids using nuclear magnetic resonance (NMR) spectroscopy. Using electron microscopy, we show that HeV nucleocapsids form herringbone-like structures typical of paramyxoviruses. We also report the crystal structure of XD of P that consists of a three-helix bundle. We study the interaction between NTAIL and XD using NMR titration experiments and provide a detailed mapping of the reciprocal binding sites. We show that the interaction is accompanied by α-helical folding of the molecular recognition element of NTAIL upon binding to a hydrophobic patch on the surface of XD. Finally, using solution NMR, we investigate the interaction between intact nucleocapsids and XD. Our results indicate that monomeric XD binds to NTAIL without triggering an additional unwinding of the nucleocapsid template. The present results provide a structural description at the atomic level of the protein-protein interactions required for transcription and replication of HeV, and the first direct observation of the interaction between the X domain of P and intact nucleocapsids in Paramyxoviridae.

【 授权许可】

CC BY