Atomic resolution description of the interaction between the nucleoprotein and phosphoprotein of Hendra virus

PLoS Pathog. 2013;9(9):e1003631. doi: 10.1371/journal.ppat.1003631. Epub 2013 Sep 26.


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, N(TAIL), 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 N(TAIL) 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 N(TAIL) 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 N(TAIL) 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 N(TAIL) 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray
  • Hendra Virus / chemistry*
  • Hendra Virus / genetics
  • Hendra Virus / metabolism
  • Humans
  • Magnetic Resonance Spectroscopy
  • Microscopy, Electron, Transmission
  • Nucleocapsid Proteins / chemistry*
  • Nucleocapsid Proteins / genetics
  • Nucleocapsid Proteins / metabolism
  • Phosphoproteins / chemistry*
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Protein Structure, Tertiary


  • Nucleocapsid Proteins
  • Phosphoproteins

Grant support

This work was carried out with financial support from the French Agence Nationale de la Recherche (ANR) through the specific programs “Physico-Chimie du Vivant” ANR-08-PCVI-0020-01, “Blanc” ANR-09-BLAN-0100 and “ASTRID” ANR-11-ASTR-003-01 (to SL), ANR JCJC ProteinDisorder (to MRJ), ANR MALZ TAUSTRUCT (to MB), the Human Frontier Science Program (long-term fellowship to RS), the Rhône-Alpes region (Ph.D. fellowship to FY), the CNRS and the Direction Générale de l'Armement (Ph.D.fellowship to DB) and the Finovi foundation (to MB and RWHR). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.