Comparative study on the biological properties of 2',5'-oligoadenylate derivatives with purified human RNase L expressed in E. coli.
An endoribonuclease, RNase L, which is activated in the presence of 2',5'-linked oligoadenylates, p(1-3)A(2'p5'A)(>2), is the terminal factor of the anti-viral action of interferon. Activation of RNase L results in inhibition of viral proliferation along with induction of apoptosis. Attempts to acquire more effective activators, 2-5A derivatives, have been made for the development of antiviral or anticancer agents. However, the ability of 2-5A derivatives to activate RNase L could not simply be compared due to the diversity of the assay methods used. We have now developed a facile method for assaying the activity of RNase L involving the use of non-fusion RNase L expressed in Escherichia coli and yeast 5S ribosomal RNA as a substrate. Using this method, several 2-5A derivative species have been revaluated. The results suggest that 2-5A molecules modified at the 8-position of the third (from the 5' terminus) adenine ring cause effective dimerization of RNase L and thus increase the ability of RNase L activation.
D000227:Adenine Nucleotides / Q000737:chemistry / Q000494:pharmacology*; D000241:Adenosine / Q000737:chemistry; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D009843:Oligoribonucleotides / Q000737:chemistry / Q000494:pharmacology*; D010957:Plasmids / Q000235:genetics; D012335:RNA, Ribosomal / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013237:Stereoisomerism
D000227:Adenine Nucleotides; D009843:Oligoribonucleotides; D012335:RNA, Ribosomal; D011994:Recombinant Proteins; C023505:2',5'-oligoadenylate; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease; D000241:Adenosine