A bulged stem-loop structure in the 3' untranslated region of the genome of the coronavirus mouse hepatitis virus is essential for replication

J Virol. 1997 Oct;71(10):7567-78. doi: 10.1128/JVI.71.10.7567-7578.1997.

Abstract

The 3' untranslated region (UTR) of the positive-sense RNA genome of the coronavirus mouse hepatitis virus (MHV) contains sequences that are necessary for the synthesis of negative-strand viral RNA as well as sequences that may be crucial for both genomic and subgenomic positive-strand RNA synthesis. We have found that the entire 3' UTR of MHV could be replaced by the 3' UTR of bovine coronavirus (BCV), which diverges overall by 31% in nucleotide sequence. This exchange between two viruses that are separated by a species barrier was carried out by targeted RNA recombination. Our results define regions of the two 3' UTRs that are functionally equivalent despite having substantial sequence substitutions, deletions, or insertions with respect to each other. More significantly, our attempts to generate an unallowed substitution of a particular portion of the BCV 3' UTR for the corresponding region of the MHV 3' UTR led to the discovery of a bulged stem-loop RNA secondary structure, adjacent to the stop codon of the nucleocapsid gene, that is essential for MHV viral RNA replication.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Composition
  • Base Sequence
  • Cattle
  • Coronavirus, Bovine / genetics
  • Genome, Viral*
  • Mice
  • Molecular Sequence Data
  • Murine hepatitis virus / chemistry
  • Murine hepatitis virus / genetics
  • Murine hepatitis virus / physiology*
  • Nucleic Acid Conformation*
  • RNA, Viral / chemistry*
  • RNA, Viral / genetics
  • Recombination, Genetic
  • Restriction Mapping
  • Transcription, Genetic
  • Virus Replication / genetics*

Substances

  • RNA, Viral