Coronavirus minus-strand RNA synthesis and effect of cycloheximide on coronavirus RNA synthesis

Journal Article · Wed Jan 01 04:00:00 EST 1986 · J. Virol.; (United States)

OSTI ID:6207536

Sawicki, S G; Sawicki, D L

The temporal sequence of coronavirus plus-strand and minus-strand RNA synthesis was determined in 17CL1 cells infected with the A59 strain of mouse hepatitis virus (MHV). MHV-induced fusion was prevented by keeping the pH of the medium below pH 6.8. This had no effect on the MHV replication cycle, but gave 5- to 10-fold-greater titers of infectious virus and delayed the detachment of cells from the monolayer which permitted viral RNA synthesis to be studied conveniently until at least 10 h postinfection. Seven species of poly(A)-containing viral RNAs were synthesized at early and late times infection, in nonequal but constant ratios. MHV minus-strand RNA synthesis was first detected at about 3 h after infection and was found exclusively in the viral replicative intermediates and was not detected in 60S single-stranded form in infected cells. Early in the replication cycle, from 45 to 65% of the (/sup 3/H)uridine pulse-labeled RF core of purified MHV replicative intermediates was in minus-strand RNA. The rate of minus-strand synthesis peaked at 5 to 6 h postinfection and then declined to about 20% of the maximum rate. The addition of cycloheximide before 3 h postinfection prevented viral RNA synthesis, whereas the addition of cycloheximide after viral RNA synthesis had begun resulted in the inhibition of viral RNA synthesis. The synthesis of both genome and subgenomic mRNAs and of viral minus strands required continued protein synthesis, and minis-strand RNA synthesis was three- to fourfold more sensitive to inhibition of cycloheximide than was plus-strand synthesis.

• Research Organization: Medical College of Ohio, Toledo (USA)
• OSTI ID: 6207536
• Journal Information: J. Virol.; (United States), Journal Name: J. Virol.; (United States) Vol. 57:1; ISSN JOVIA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
AZINES
BIOLOGICAL EFFECTS
BIOSYNTHESIS
CYCLOHEXIMIDE
DRUGS
FUNGICIDES
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
LABELLED COMPOUNDS
MICROORGANISMS
MOLECULAR BIOLOGY
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PESTICIDES
PYRIMIDINES
RIBOSIDES
RNA
SYNTHESIS
TRITIUM COMPOUNDS
URACILS
URIDINE
VIRUSES