In vitro translation of polyadenylated RNA isolated from control and interferon-treated human promyelocytic HL-60 leukemic cells.
Polyadenylated RNA has been isolated from control and interferon-treated HL-60 cells by centrifugation through cesium chloride and oligo(dT)-cellulose column chromatography. The affinity column-purified RNA is poorly translated in the mRNA-dependent rabbit reticulocyte lysates but is an excellent template for in vitro protein synthesis using the wheat germ cell extracts. The discrepancy in the efficiency of HL-60 mRNA utilization in the two commonly used cell-free protein synthesizing systems is attributable to an inhibitory component present in the polyadenylated RNA. This contaminant is most likely double-stranded RNA based on (i) the ability of 2-aminopurine (3-5 mM) or high concentrations of penicillium chrysogenum double-stranded RNA (10-15 micrograms/ml) to overcome the inhibition exerted by the component, and (ii) the ability of the component to promote the enzymatic conversion of ATP into 2-5A by the highly purified rabbit reticulocyte 2-5A synthetase.
T J
TJ
Borelli
J M
JM
Wu
D015088:2',5'-Oligoadenylate Synthetase / Q000378:metabolism; D000818:Animals; D002474:Cell-Free System; D002478:Cells, Cultured; D006801:Humans; D066298:In Vitro Techniques; D007370:Interferon Type I / Q000494:pharmacology*; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000096:biosynthesis; D011061:Poly A / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects*; D012313:RNA / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000378:metabolism; D012333:RNA, Messenger; D012334:RNA, Neoplasm / Q000378:metabolism; D011817:Rabbits
D007370:Interferon Type I; D009363:Neoplasm Proteins; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D011061:Poly A; D012313:RNA; D015088:2',5'-Oligoadenylate Synthetase