title: Generation and propagation of recombinant mumps viruses exhibiting an additional U residue in the homopolymeric U tract of the F gene-end signal.
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.
authors.affiliation: Food and Drug Administration (FDA), Center for Biologics Evaluation and Research (CBER), OVRR, DVP, 10903 New Hampshire Avenue, Bldg 52/72, Silver Spring, MD, 20993, USA, christian.sauder@fda.hhs.gov.
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Sauder
authors.forename: Laurie
authors.initials: L
authors.lastname: Ngo
authors.forename: Vahan
authors.initials: V
authors.lastname: Simonyan
authors.forename: Yu
authors.initials: Y
authors.lastname: Cong
authors.forename: Cheryl
authors.initials: C
authors.lastname: Zhang
authors.forename: Malen
authors.initials: M
authors.lastname: Link
authors.forename: Tahir
authors.initials: T
authors.lastname: Malik
authors.forename: Steven A
authors.initials: SA
authors.lastname: Rubin
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D009109:Mumps virus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D016254:Mutagenesis, Insertional; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics; D017422:Sequence Analysis, DNA; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D010948:Viral Plaque Assay; D014774:Virulence; D014779:Virus Replication
chemical_list: D050505:Mutant Proteins; D012367:RNA, Viral; D014760:Viral Fusion Proteins