Two murine coronavirus genes suffice for viral RNA synthesis

Hwang Kim Kyongmin Hwang Kim, Shinji Makino

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

We identified two mouse hepatitis virus (MHV) genes that suffice for MHV RNA synthesis by using an MHV-JHM-derived defective interfering (DI) RNA, DIssA. DIssA is a naturally occurring self-replicating DI RNA with nearly intact genes 1 and 7. DIssA interferes with most MHV-JHM-specific RNA synthesis, except for synthesis of mRNA 7, which encodes N protein; mRNA 7 synthesis is not inhibited by DIssA. Coinfection of MHV-JHM containing DIssA DI particles and an MHV-A59 RNA temperature-sensitive mutant followed by subsequent passage of virus at the permissive temperature resulted in elimination of most of the MHV-JHM helper virus. Analysis of intracellular RNAs at the nonpermissive temperature demonstrated efficient synthesis of DIssA and mRNA 7 but not of the helper virus mRNAs. Oligonucleotide fingerprinting analysis demonstrated that the structure of mRNA 7 was MHV- JHM specific and therefore must have been synthesized from the DIssA template RNA. Sequence analysis revealed that DIssA lacks a slightly heterogeneous sequence, which is found in wild-type MHV from the 3' one-third of gene 2-1 to the 3' end of gene 6. Northern (RNA) blot analysis of intracellular RNA species and virus-specific protein analysis confirmed the sequence data. Replication and transcription of another MHV DI RNA were supported in DIssA- replicating cells. Because the products of genes 2 and 2-1 are not essential for MHV replication, we concluded that expression of gene I proteins and N protein was sufficient for MHV RNA replication and transcription.

Original languageEnglish (US)
Pages (from-to)2313-2321
Number of pages9
JournalJournal of Virology
Volume69
Issue number4
StatePublished - 1995
Externally publishedYes

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Murine coronavirus
Murine hepatitis virus
Coronavirus
Viral RNA
RNA
synthesis
Genes
genes
Messenger RNA
Helper Viruses
viruses
Virus Replication
Temperature
sequence analysis
proteins
transcription (genetics)
Defective Viruses
temperature
Proteins
RNA Viruses

ASJC Scopus subject areas

  • Immunology

Cite this

Two murine coronavirus genes suffice for viral RNA synthesis. / Kyongmin Hwang Kim, Hwang Kim; Makino, Shinji.

In: Journal of Virology, Vol. 69, No. 4, 1995, p. 2313-2321.

Research output: Contribution to journalArticle

Kyongmin Hwang Kim, HK & Makino, S 1995, 'Two murine coronavirus genes suffice for viral RNA synthesis', Journal of Virology, vol. 69, no. 4, pp. 2313-2321.
Kyongmin Hwang Kim, Hwang Kim ; Makino, Shinji. / Two murine coronavirus genes suffice for viral RNA synthesis. In: Journal of Virology. 1995 ; Vol. 69, No. 4. pp. 2313-2321.
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