Nature and mode of action of vaccinia virus products that block activation of the interferon-mediated ppp(A2'p)nA-synthetase.
In this report it has been shown that inhibition of the 2-5A synthetase in IFN-treated, vaccinia virus-infected mouse L and human HeLa S3 cells is related to specific viral functions. This inhibition occurs concomitantly with degradation of ATP and with dephosphorylation of ppp(A2'p)nA. At least two viral-mediated enzyme activities are thought to be involved in this process, an ATPase and a phosphatase. The ATPase activity was established after determining the extent of hydrolysis of ATP, the nature of 2-5A, and the relative abundance of the different oligomers. Cytoplasmic cell extracts and purified vaccinia virions were bound to poly (I):(C) agarose, incubated with [3H]ATP, [alpha-32P]ATP, or [gamma-32P]ATP, and the extent of hydrolysis of ATP was determined by TLC. Authentic 2-5A and the relative abundance of the various oligomers were characterized by enzymatic and alkali treatments and identification by TLC and HPLC analysis. The phosphatase activity was measured by TLC after determining the degree of dephosphorylation of 2-5A from the extent of labeling at the 5'-OH termini with [gamma-32P]ATP and polynucleotide kinase. While free 5'-OH termini were not observed in oligomers synthesized with bound poly (I):(C) agarose enzyme fractions from IFN-treated, uninfected cells, a strong phosphorylation was found in oligomers from IFN-treated, infected cells. These findings suggest that it is the contribution of these viral enzyme activities that renders vaccinia virus resistant to interferons.
D015088:2',5'-Oligoadenylate Synthetase / Q000037:antagonists & inhibitors*; D000227:Adenine Nucleotides / Q000096:biosynthesis / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D004790:Enzyme Induction; D006367:HeLa Cells; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology*; D007739:L Cells; D051379:Mice; D009841:Oligonucleotides / Q000378:metabolism; D009843:Oligoribonucleotides / Q000096:biosynthesis / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010766:Phosphorylation; D014616:Vaccinia virus / Q000201:enzymology* / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis
D000227:Adenine Nucleotides; D007370:Interferon Type I; D009841:Oligonucleotides; D009843:Oligoribonucleotides; D014764:Viral Proteins; C023505:2',5'-oligoadenylate; D000255:Adenosine Triphosphate; D015088:2',5'-Oligoadenylate Synthetase; D010744:Phosphoric Monoester Hydrolases; D000251:Adenosine Triphosphatases