Activation of 2',5'-oligo(A) polymerase and protein kinase of interferon-treated HeLa cells by 2'-O-methylated poly (inosinic acid) . poly(cytidylic acid), Correlations with interferon-inducing activity.
An oligonucleotide polymerase and a protein kinase which require double-stranded RNA (dsRNA) for activation are induced in HeLa cells by human fibroblast interferon. The polymerase synthesizes a series of oligonucleotides from ATP, whereas the kinase phosphorylates a polypeptide of Mr = 72,000 and the alpha subunit of initiation factor eIF-2. Partially or fully 2'-O-methylated derivatives of poly(inosinic acid) . poly(cytidylic acid) (rIn . rCn) were used to determine the structural requirements of dsRNA in the activation of these two enzymes. While fully methylated polymers failed to activate either enzyme, partially methylated polymers activated the enzymes in specific manners. The activation of the kinase by the rIn . rCn analogues was affected more severely by the level of methylation than was the activation of the polymerase. Moreover, fully methylated analogues blocked the activation of the kinase by rIn . rCn but not the activation of the polymerase. These observations are consistent with a biphasic model for enzyme activation similar to that proposed for interferon induction, which required the recognition of a relatively small region of rIn . rCn as the last step. Differences in the activation of the polymerase and kinase are explicable on the basis of the polymerase requirement for a smaller recognition region of the rIn . rCn duplex than the kinase. Dependence of polymerase activation on the level of methylation shows striking similarities with the interferon inducing activities of these analogues, suggesting a possible relationship between polymerase activation and interferon induction.
D015088:2',5'-Oligoadenylate Synthetase; D000227:Adenine Nucleotides / Q000096:biosynthesis; D000255:Adenosine Triphosphate / Q000378:metabolism; D004789:Enzyme Activation; D006367:HeLa Cells / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D007372:Interferons / Q000096:biosynthesis / Q000494:pharmacology*; D008745:Methylation; D009841:Oligonucleotides / Q000096:biosynthesis; D009843:Oligoribonucleotides / Q000096:biosynthesis; D011070:Poly I-C / Q000494:pharmacology*; D011062:Polynucleotide Adenylyltransferase / Q000378:metabolism; D011118:Polynucleotide Ligases / Q000096:biosynthesis*; D011494:Protein Kinases / Q000096:biosynthesis* / Q000378:metabolism
D000227:Adenine Nucleotides; D009841:Oligonucleotides; D009843:Oligoribonucleotides; D000255:Adenosine Triphosphate; D007372:Interferons; D011494:Protein Kinases; D011062:Polynucleotide Adenylyltransferase; D015088:2',5'-Oligoadenylate Synthetase; D011118:Polynucleotide Ligases; D011070:Poly I-C