Structural and functional characterization of the mumps virus phosphoprotein

J Virol. 2013 Jul;87(13):7558-68. doi: 10.1128/JVI.00653-13. Epub 2013 May 1.

Abstract

The phosphoprotein (P) is virally encoded by the Rhabdoviridae and Paramyxoviridae in the order Mononegavirales. P is a self-associated oligomer and forms complexes with the large viral polymerase protein (L), the nucleocapsid protein (N), and the assembled nucleocapsid. P from different viruses has shown structural diversities even though their essential functions are the same. We systematically mapped the domains in mumps virus (MuV) P and investigated their interactions with nucleocapsid-like particles (NLPs). Similar to other P proteins, MuV P contains N-terminal, central, and C-terminal domains with flexible linkers between neighboring domains. By pulldown assays, we discovered that in addition to the previously proposed nucleocapsid binding domain (residues 343 to 391), the N-terminal region of MuV P (residues 1 to 194) could also bind NLPs. Further analysis of binding kinetics was conducted using surface plasmon resonance. This is the first observation that both the N- and C-terminal regions of a negative-strand RNA virus P are involved in binding the nucleocapsid. In addition, we defined the oligomerization domain (POD) of MuV P as residues 213 to 277 and determined its crystal structure. The tetrameric MuV POD is formed by one pair of long parallel α-helices with another pair in opposite orientation. Unlike the parallel orientation of each α-helix in the tetramer of Sendai virus POD, this represents a novel orientation of a POD where both the N- and the C-terminal domains are at either end of the tetramer. This is consistent with the observation that both the N- and the C-terminal domains are involved in binding the nucleocapsid.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computational Biology
  • Crystallization
  • Escherichia coli
  • Kinetics
  • Models, Molecular*
  • Mumps virus / genetics*
  • Nucleocapsid Proteins / metabolism
  • Phosphoproteins / chemistry*
  • Phosphoproteins / genetics*
  • Plasmids / genetics
  • Protein Conformation*
  • Protein Structure, Tertiary
  • Surface Plasmon Resonance
  • X-Ray Diffraction

Substances

  • Nucleocapsid Proteins
  • Phosphoproteins