Nitric oxide inhibition of coxsackievirus replication in vitro.
Nitric oxide is a radical molecule with antibacterial, -parasitic, and -viral properties. We investigated the mechanism of NO inhibition of Coxsackievirus B3 (CVB3) replication in vitro by determining the effect of NO upon a single replicative cycle of CVB3 grown in HeLa cells. Transfection of inducible NO synthase cDNA into HeLa cells reduces the number of viral particles produced during a single cycle of growth. Similarly, a noncytotoxic concentration of the NO donor S-nitroso-amino-penicillamine reduces the number of viral particles in a dose-dependent manner. To explore the mechanisms by which NO exerts its antiviral effect, we assayed the attachment, replication, and translation steps of the CVB3 life cycle. NO does not affect the attachment of CVB3 to HeLa cells. However, NO inhibits CVB3 RNA synthesis, as shown by a [3H]uridine incorporation assay, reverse transcription-PCR, and Northern analysis. In addition, NO inhibits CVB3 protein synthesis, as shown by [35S]methionine protein labeling and Western blot analysis of infected cells. Thus, NO inhibits CVB3 replication in part by inhibiting viral RNA synthesis by an unknown mechanism.
Zaragoza
C
C
Division of Cardiology, Department of Medicine, School of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Ocampo
C J
CJ
Saura
M
M
McMillan
A
A
Lowenstein
C J
CJ
D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004770:Enterovirus / Q000187:drug effects* / Q000254:growth & development; D006367:HeLa Cells; D006801:Humans; D009569:Nitric Oxide / Q000494:pharmacology*; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism; D010397:Penicillanic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D012367:RNA, Viral / Q000096:biosynthesis; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication / Q000187:drug effects*
D012367:RNA, Viral; D014764:Viral Proteins; D009569:Nitric Oxide; D010397:Penicillanic Acid; D019001:Nitric Oxide Synthase; C004408:aminopenicillanic acid