Nitric oxide inhibits the adenovirus proteinase in vitro and viral infectivity in vivo.
Nitric oxide (NO) is an antiviral effector of the innate immune system, but few of the viral targets of NO have been identified. We now show that NO inhibits adenovirus replication by targeting the adenovirus proteinase (AVP). NO generated from diethylamine NONOate (DEA-NONOate) or spermine NONOate (Sp-NONOate) inhibited the AVP. Inhibition was reversible with dithiothreitol. The equilibrium dissociation constant for reversible binding to the AVP by Sp-NONOate, or Ki, was 0.47 mM, and the first-order rate constant for irreversible inhibition of the AVP by Sp-NONOate, or ki, was 0.0036 s(-1). Two hallmarks of a successful adenovirus infection were abolished by the NO donors: the appearance of E1A protein and the cleavage of cytokeratin 18 by AVP. Treatment of infectious virus by DEA-NONOate dramatically decreased viral infectivity. These data suggest that NO may be a useful antiviral agent against viruses encoding a cysteine proteinase and in particular may be an antiadenovirus agent.
Cao
Wangsen
W
Division of Cardiology, Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Baniecki
Mary Lynn
ML
McGrath
William J
WJ
Bao
Clare
C
Deming
Clayton B
CB
Rade
Jeffrey J
JJ
Lowenstein
Charles J
CJ
Mangel
Walter F
WF
D000256:Adenoviridae / Q000187:drug effects* / Q000201:enzymology*; D000257:Adenoviridae Infections / Q000188:drug therapy; D000998:Antiviral Agents / Q000494:pharmacology* / Q000627:therapeutic use; D003546:Cysteine Endopeptidases / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D004229:Dithiothreitol / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D006834:Hydrazines / Q000494:pharmacology; D008954:Models, Biological; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000494:pharmacology* / Q000627:therapeutic use; D009589:Nitrogen Oxides; D010446:Peptide Fragments / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism
D000998:Antiviral Agents; D015853:Cysteine Proteinase Inhibitors; D006834:Hydrazines; D020030:Nitric Oxide Donors; D009589:Nitrogen Oxides; D010446:Peptide Fragments; D014764:Viral Proteins; C105093:glycyl-valyl-glutaminyl-seryl-leucyl-lysyl-arginyl-arginyl-arginyl-cysteinyl-phenylalanine; D009569:Nitric Oxide; C084012:1,1-diethyl-2-hydroxy-2-nitrosohydrazine; D003546:Cysteine Endopeptidases; D004229:Dithiothreitol