Regulation of measles virus gene expression by P protein coiled-coil properties

Sci Adv. 2019 May 8;5(5):eaaw3702. doi: 10.1126/sciadv.aaw3702. eCollection 2019 May.

Abstract

The polymerase of negative-stranded RNA viruses consists of the large protein (L) and the phosphoprotein (P), the latter serving both as a chaperon and a cofactor for L. We mapped within measles virus (MeV) P the regions responsible for binding and stabilizing L and showed that the coiled-coil multimerization domain (MD) of P is required for gene expression. MeV MD is kinked as a result of the presence of a stammer. Both restoration of the heptad regularity and displacement of the stammer strongly decrease or abrogate activity in a minigenome assay. By contrast, P activity is rather tolerant of substitutions within the stammer. Single substitutions at the "a" or "d" hydrophobic anchor positions with residues of variable hydrophobicity revealed that P functionality requires a narrow range of cohesiveness of its MD. Results collectively indicate that, beyond merely ensuring P oligomerization, the MD finely tunes viral gene expression through its cohesiveness.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Gene Expression Regulation, Viral*
  • HEK293 Cells
  • Humans
  • Measles virus / metabolism*
  • Molecular Dynamics Simulation
  • Mutagenesis
  • Paramyxoviridae / metabolism
  • Phosphoproteins / antagonists & inhibitors
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism*
  • Protein Conformation, alpha-Helical
  • Protein Domains
  • Protein Folding
  • Protein Multimerization
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Viral Proteins / antagonists & inhibitors
  • Viral Proteins / chemistry
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • P protein, Sendai virus
  • Phosphoproteins
  • RNA, Small Interfering
  • Recombinant Proteins
  • Viral Proteins