Segment-specific terminal sequences of Bunyamwera bunyavirus regulate genome replication

J N Barr, Richard Michael Elliott, E F Dunn, G W Wertz

Research output: Contribution to journalArticlepeer-review

Abstract

Bunyamwera virus (BUNV) is the prototype of both the Orthobunyavirus genus and the Bunyaviridae family of segmented negative sense RNA viruses. The tripartite BUNV genome consists of small (S), medium (M), and large (L) segments that are transcribed to give a single mRNA and replicated to generate an antigenome that is the template for synthesis of further genomic RNA strands. We modified an existing cDNA-derived RNA synthesis system to allow identification of BUNV RNA replication and transcription products by direct metabolic labeling. Direct RNA analysis allowed us to distinguish between template activities that affected either RNA replication or mRNA transcription, an ability that was not possible using previous reporter gene expression assays. We generated genome analogs containing the entire nontranslated terminal sequences of the S, M, and L BUNV segments surrounding a common sequence. Analysis of RNAs synthesized from these templates revealed that the relative abilities of BUNV segments to perform RNA replication was M > L > S. Exchange of segment-specific terminal nucleotides identified a 12-nt region located within both the 3' and 5' termini of the M segment that correlated with its high replication ability. (C) 2003 Elsevier Science (USA). All rights reserved.

Original languageEnglish
Pages (from-to)326-338
Number of pages13
JournalVirology
Volume311
Issue number2
DOIs
Publication statusPublished - 5 Jul 2003

Keywords

  • Bunyamwera virus
  • Bunyaviridae
  • genome replication
  • mRNA transcription
  • unique segment terminal sequences
  • VESICULAR STOMATITIS-VIRUS
  • INFLUENZA-A VIRUS
  • NONSTRUCTURAL PROTEIN NSS
  • VIRAL MESSENGER-RNAS
  • VALLEY FEVER VIRUS
  • LA CROSSE VIRUS
  • UUKUNIEMI VIRUS
  • NUCLEOTIDE-SEQUENCE
  • FAMILY BUNYAVIRIDAE
  • MUTATIONAL ANALYSIS

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