Effect of actinomycin D on simian rotavirus (SA11) replication in cell culture.
Rotaviruses are the major cause of viral diarrhea in humans and animals. Actinomycin D (Act D) is an antibiotic that intercalates DNA and therefore inhibits DNA-dependent transcription. The current study was carried out to assess the influence of Act D on the replication of simian rotavirus (SA11) in cell culture. Virus-infected MA-104 cell cultures were studied in the presence of Act D at concentrations of 1.25 and 2.5 microg/ml. Treatment of rotavirus-infected cells with 2.5 microg/ml Act D 48 h post-infection reduced the cytoplasmic metachromasia after staining with acridine orange by 25%. Viral RNA labeled with 3H-uridine in the presence of the drug was separated by polyacrylamide gel electrophoresis. Viral RNA replication was not affected by Act D, but increased 3H-uridine uptake was demonstrable by infected cells in the presence of the drug. This possibly was due to the inhibition of cellular RNA synthesis by Act D, which thus enhances incorporation of the radionuclide into the viral RNA. Act D reduced the number of infected cells presenting virus-specific fluorescence 48 h post-infection by more than 50%. These data suggest that Act D may have complexed with viral RNA and prevented newly synthesized mRNA from being translated, but may not have prevented early replication.
Stefanelli
C C
CC
Laboratório de Virologia, Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, PR, Brasil.
Castilho
J G
JG
Botelho
M V J
MV
Linhares
R E C
RE
Nozawa
C M
CM
D000818:Animals; D018929:Cell Culture Techniques; D003609:Dactinomycin / Q000494:pharmacology*; D008253:Macaca mulatta; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology*; D012367:RNA, Viral / Q000187:drug effects*; D012401:Rotavirus / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects* / Q000502:physiology
D019384:Nucleic Acid Synthesis Inhibitors; D012367:RNA, Viral; D003609:Dactinomycin