Generation and propagation of recombinant mumps viruses exhibiting an additional U residue in the homopolymeric U tract of the F gene-end signal

Virus Genes. 2015 Aug;51(1):12-24. doi: 10.1007/s11262-015-1204-y. Epub 2015 May 12.

Abstract

As a member of the family paramyxoviridae, subfamily paramyxovirinae, the genome of mumps virus (MuV) is postulated to be polyhexameric in length in order to be able to replicate efficiently. While all natural MuV strains sequenced so far obey to this "rule of six," we describe here the isolation of recombinant MuVs that appeared to contain an additional U residue in the homopolymeric tract of the F gene-end signal, resulting in a genome length of 6n + 1. Sequencing of several plaque-purified viruses from these preparations did not reveal the existence of length-correcting mutations, suggesting that they are violators of the rule of six. Employing high-throughput sequencing technology, we provide evidence that the insertion of an additional U residue is mainly the result of the rescue system used that relies on T7 RNA polymerase. Limited in vitro and in vivo testing of the viruses did not reveal any significant impact of the longer genome on virus replication or virulence, suggesting that the rule of six is not a strict requirement for MuV replication.

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • Mumps virus / genetics
  • Mumps virus / isolation & purification
  • Mumps virus / physiology*
  • Mutagenesis, Insertional*
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism*
  • RNA, Viral / genetics
  • Sequence Analysis, DNA
  • Vero Cells
  • Viral Fusion Proteins / genetics
  • Viral Fusion Proteins / metabolism*
  • Viral Plaque Assay
  • Virulence
  • Virus Replication*

Substances

  • Mutant Proteins
  • RNA, Viral
  • Viral Fusion Proteins