TY - JOUR
T1 - Nitric oxide inhibition of coxsackievirus replication in vitro
AU - Zaragoza, Carlos
AU - Ocampo, Christopher J.
AU - Saura, Marta
AU - McMillan, Audrey
AU - Lowenstein, Charles J.
PY - 1997/10/1
Y1 - 1997/10/1
N2 - 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 vital 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.
AB - 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 vital 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.
KW - Autoimmune
KW - Enterovirus
KW - Myocarditis
KW - Nitric oxide synthase
KW - Nuclear factor kappa B
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U2 - 10.1172/JCI119702
DO - 10.1172/JCI119702
M3 - Article
C2 - 9312175
AN - SCOPUS:0030931643
SN - 0021-9738
VL - 100
SP - 1760
EP - 1767
JO - Journal of Clinical Investigation
JF - Journal of Clinical Investigation
IS - 7
ER -