2',5'-Oligo(A)-activated endoribonuclease. Tissue distribution and characterization with a binding assay.
We have measured the binding of highly radioactive (2'-5')pppA4-5[32P]cytidine 3',5'-diphosphate to human, mouse, and rabbit cell and tissue extracts. A binding activity for this oligonucleotide is present in all extracts examined and high levels of this activity are found in some cells of lymphoid origin. In particular, the mouse thymoma cell line W7 has about 10 times higher binding activity than other cell lines. The oligonucleotide is bound with a single high affinity constant, as shown by Scatchard plot analyses. Cell extracts fractionated by centrifugation on glycerol gradients show a single peak of binding activity, which co-sediments with (2'-5')oligo(A)-dependent endoribonuclease (RNase L). This enzyme apparently binds the oligonucleotide, as shown by experiments with an analog of (2'-5')oligo(A) which competes in the binding and inhibits the RNase L. This endonuclease cannot be directly assayed in unfractionated cell or tissue extracts with high levels of other nuclease activities. The binding assay for the oligonucleotide may provide an estimate of RNase L levels in such cell and tissue extracts.
D000227:Adenine Nucleotides / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D004722:Endoribonucleases; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D007739:L Cells / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008214:Lymphocytes / Q000201:enzymology; D051379:Mice; D008970:Molecular Weight; D009841:Oligonucleotides / Q000494:pharmacology*; D009843:Oligoribonucleotides / Q000494:pharmacology*; D011485:Protein Binding; D012156:Reticulocytes / Q000201:enzymology; D012260:Ribonucleases / Q000378:metabolism*; D013045:Species Specificity; D013154:Spleen / Q000201:enzymology; D014018:Tissue Distribution
D000227:Adenine Nucleotides; D009841:Oligonucleotides; D009843:Oligoribonucleotides; C023505:2',5'-oligoadenylate; D004722:Endoribonucleases; D012260:Ribonucleases; C031956:2-5A-dependent ribonuclease