Interferons, double-stranded RNA, and RNA degradation. Isolation and characterization of homogeneous human (2'-5')(a)n synthetase.
(2'-5')(A)n synthetase is one of the mediators of interferon action. If activated by double-stranded RNA it converts ATP into pyrophosphate and (2'-5')(A)n. In turn, (2'-5')(A)n activates a latent endoribonuclease (RNase L) which cleaves single-stranded RNA. We report here the isolation and characterization of a homogeneous human (2'-5')(A)n synthetase. The enzyme was purified from interferon-treated HeLA S3 cells by chromatography of a ribosomal salt wash fraction on DEAE-cellulose, poly(I) . poly(C) agarose, and CM-cellulose. The purified (2'-5')(A)n synthetase can convert over 90% of ATP into (2'-5')(A)n. The enzyme is unstable but can be stabilized by certain nonionic detergents (e.g. Triton X-100). Its apparent Mr = 100,000, as determined by gel electrophoresis in sodium dodecyl sulfate, and about 80,000, as determined by centrifugation through a glycerol gradient. The human (2'-5')(A)n synthetase is similar to the corresponding enzyme from mouse Ehrlich ascites tumor cells, but differs from the latter in size (100,000 versus 105,000 daltons) and in ionic conditions required for maximal activity.
D015088:2',5'-Oligoadenylate Synthetase; D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000201:enzymology; D002846:Chromatography, Affinity; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007372:Interferons / Q000494:pharmacology*; D051379:Mice; D011070:Poly I-C; D011118:Polynucleotide Ligases / Q000302:isolation & purification* / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000494:pharmacology*
D012330:RNA, Double-Stranded; D007372:Interferons; D015088:2',5'-Oligoadenylate Synthetase; D011118:Polynucleotide Ligases; D011070:Poly I-C