Temporal regulation of bovine coronavirus RNA synthesis

James G. Keck, Brenda Hogue, David A. Brian, Michael M C Lai

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The structure and synthesis of bovine coronavirus (BCV)-specific intracellular RNA were studied. A genome-size RNA and seven subgenomic RNAs with molecular weights of approximately 3.3 × 106, 3.1 × 106, 2.6 × 106, 1.1 × 106, 1.0 × 106, 0.8 × 106 and 0.6 × 106 were detected. Comparisons of BCV intracellular RNAs with those of mouse hepatitis virus (MHV) demonstrated the presence of an additional RNA for BCV, species 2a, of 3.1 × 106 daltons. BCV RNAs contain a nested-set structure similar to that of other coronaviruses. This nested-set structure would suggest that the new RNA has a capacity to encode a protein of approximately 430 amino acids. Kinetic studies demonstrated that the synthesis of subgenomic mRNAs and genomic RNA are differentially regulated. At 4 to 8 h post-infection (p.i.), subgenomic RNAs are synthesized at a maximal rate and represent greater than 90% of the total viral RNA synthesized, whereas genome-size RNA accounts for only 7%. Later in infection, at 70 to 72 h p.i., genome-size RNA is much more abundant and accounts for 88% of total RNA synthesized. Immunoprecipitations of [35S]methionine-pulse-labeled viral proteins demonstrated that viral protein synthesis occurs early in the infection, concurrent with the peak of viral subgenomic RNA synthesis. Western blot analysis suggests that these proteins are stable since the proteins are present at high level as late as 70 to 72 h p.i. The kinetics of production of virus particles coincides with the synthesis of genomic RNA. These studies thus indicate that there is a differential temporal regulation of the synthesis of genomic RNA and subgenomic mRNAs, and that the synthesis of genomic RNA is the rate-limiting step regulating the production of virus particles.

Original languageEnglish (US)
Pages (from-to)343-356
Number of pages14
JournalVirus Research
Volume9
Issue number4
DOIs
StatePublished - 1988
Externally publishedYes

Fingerprint

Bovine Coronavirus
RNA
Genome Size
Infection
Viral RNA
Viral Proteins
Virion
Murine hepatitis virus
Messenger RNA
Coronavirus
Proteins

Keywords

  • Bovine coronavirus
  • Coronavirus
  • RNA

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

Cite this

Temporal regulation of bovine coronavirus RNA synthesis. / Keck, James G.; Hogue, Brenda; Brian, David A.; Lai, Michael M C.

In: Virus Research, Vol. 9, No. 4, 1988, p. 343-356.

Research output: Contribution to journalArticle

Keck, James G. ; Hogue, Brenda ; Brian, David A. ; Lai, Michael M C. / Temporal regulation of bovine coronavirus RNA synthesis. In: Virus Research. 1988 ; Vol. 9, No. 4. pp. 343-356.
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AB - The structure and synthesis of bovine coronavirus (BCV)-specific intracellular RNA were studied. A genome-size RNA and seven subgenomic RNAs with molecular weights of approximately 3.3 × 106, 3.1 × 106, 2.6 × 106, 1.1 × 106, 1.0 × 106, 0.8 × 106 and 0.6 × 106 were detected. Comparisons of BCV intracellular RNAs with those of mouse hepatitis virus (MHV) demonstrated the presence of an additional RNA for BCV, species 2a, of 3.1 × 106 daltons. BCV RNAs contain a nested-set structure similar to that of other coronaviruses. This nested-set structure would suggest that the new RNA has a capacity to encode a protein of approximately 430 amino acids. Kinetic studies demonstrated that the synthesis of subgenomic mRNAs and genomic RNA are differentially regulated. At 4 to 8 h post-infection (p.i.), subgenomic RNAs are synthesized at a maximal rate and represent greater than 90% of the total viral RNA synthesized, whereas genome-size RNA accounts for only 7%. Later in infection, at 70 to 72 h p.i., genome-size RNA is much more abundant and accounts for 88% of total RNA synthesized. Immunoprecipitations of [35S]methionine-pulse-labeled viral proteins demonstrated that viral protein synthesis occurs early in the infection, concurrent with the peak of viral subgenomic RNA synthesis. Western blot analysis suggests that these proteins are stable since the proteins are present at high level as late as 70 to 72 h p.i. The kinetics of production of virus particles coincides with the synthesis of genomic RNA. These studies thus indicate that there is a differential temporal regulation of the synthesis of genomic RNA and subgenomic mRNAs, and that the synthesis of genomic RNA is the rate-limiting step regulating the production of virus particles.

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