Biochemical and structural studies of the oligomerization domain of the Nipah virus phosphoprotein: evidence for an elongated coiled-coil homotrimer.
Nipah virus (NiV) is a recently emerged severe human pathogen that belongs to the Henipavirus genus within the Paramyxoviridae family. The NiV genome is encapsidated by the nucleoprotein (N) within a helical nucleocapsid that is the substrate used by the polymerase for transcription and replication. The polymerase is recruited onto the nucleocapsid via its cofactor, the phosphoprotein (P). The NiV P protein has a modular organization, with alternating disordered and ordered domains. Among these latter, is the P multimerization domain (PMD) that was predicted to adopt a coiled-coil conformation. Using both biochemical and biophysical approaches, we show that NiV PMD forms a highly stable and elongated coiled-coil trimer, a finding in striking contrast with respect to the PMDs of Paramyxoviridae members investigated so far that were all found to tetramerize. The present results therefore represent the first report of a paramyxoviral P protein forming trimers.
CNRS and Aix-Marseille Université, Architecture et Fonction des Macromolécules Biologiques (AFMB), UMR 7257, 13288 Marseille, France.
David
D
Blocquel
Matilde
M
Beltrandi
Jenny
J
Erales
Pascale
P
Barbier
Sonia
S
Longhi
D006801:Humans; D045405:Nipah Virus / Q000737:chemistry* / Q000502:physiology*; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D055503:Protein Multimerization; D053838:Scattering, Small Angle; D014461:Ultracentrifugation; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism*
C543301:P protein, Nipah virus; D010750:Phosphoproteins; D014764:Viral Proteins
2,2,2-trifluoroethanol; Analytical ultracentrifugation; CD; Coiled-coil; Cross-linking; GF; HeV; Hendra virus; Henipavirus; Homotrimer; IMAC; L; MALDI-TOF; MRE; MeV; MuV; N; NMR; NiV; Nipah virus; OD; P; P C-terminal domain; P N-terminal domain; P multimerization domain; PCR; PCT; PMD; PNT; Phosphoprotein; R(S); R(g); RDV; RSV; RV; Rinderpest virus; SAB; SAXS; SDS-PAGE; SeV; Sendai virus; Small-angle X-ray scattering; Stokes radius; Suberic acid bis (N-hydroxy-succinimide ester); T(m); TFE; VSV; X domain of P; XD; circular dichroism; gel filtration; immobilized metal affinity chromatography; large protein; matrix-assisted laser desorption ionization/time of flight; mean ellipticity values per residue; measles virus; melting temperature; mumps virus; nuclear magnetic resonance; nucleoprotein; optical density; phosphoprotein; polymerase chain reaction; rabies virus; radius of gyration; respiratory syncytial virus; small angle X-ray scattering; sodium dodecyl sulphate polyacrylamide electrophoresis; vesicular stomatitis virus