Plant oligoadenylates: enzymatic synthesis, isolation, and biological activities.
An enzyme that converts [3H, 32P]ATP, with a 3H:32P ratio of 1:1, to oligoadenylates with the same 3H:32P ratio was increased in plants following treatment with human leukocyte interferon or plant antiviral factor or inoculation with tobacco mosaic virus. The enzyme was extracted from tobacco leaves, callus tissue cultures, or cell suspension cultures. The enzyme, a putative plant oligoadenylate synthetase, was immobilized on poly(rI) . poly(rC)-agarose columns and converted ATP into plant oligoadenylates. These oligoadenylates were displaced from DEAE-cellulose columns with 350 mM KCl buffer, dialyzed, and further purified by high-performance liquid chromatography (HPLC) and DEAE-cellulose gradient chromatography. In all steps of purification, the ratio of 3H:32P in the oligoadenylates remained 1:1. The plant oligoadenylates isolated by displacement with 350 mM KCl had a molecular weight greater than 1000. The plant oligoadenylates had charges of 5- and 6-. HPLC resolved five peaks, three of which inhibited protein synthesis in reticulocyte and wheat germ systems. Partial structural elucidation of the plant oligoadenylates has been determined by enzymatic and chemical treatments. An adenylate with a 3',5'-phosphodiester and/or a pyrophosphoryl linkage with either 3'- or 5'-terminal phosphates is postulated on the basis of treatment of the oligoadenylates with T2 RNase, snake venom phosphodiesterase, and bacterial alkaline phosphatase and acid and alkaline hydrolyses. The plant oligoadenylates at 8 X 10(-7) M inhibit protein synthesis by 75% in lysates from rabbit reticulocytes and 45% in wheat germ cell-free systems.(ABSTRACT TRUNCATED AT 250 WORDS)
D015088:2',5'-Oligoadenylate Synthetase / Q000235:genetics; D000227:Adenine Nucleotides / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D004722:Endoribonucleases / Q000097:blood; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology; D007700:Kinetics; D009841:Oligonucleotides / Q000096:biosynthesis*; D009843:Oligoribonucleotides / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D010761:Phosphorus Radioisotopes; D010944:Plants / Q000187:drug effects / Q000201:enzymology*; D010947:Plants, Toxic; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology; D014026:Nicotiana / Q000201:enzymology; D014316:Tritium
D000227:Adenine Nucleotides; D007370:Interferon Type I; D009841:Oligonucleotides; D009843:Oligoribonucleotides; D010761:Phosphorus Radioisotopes; D014316:Tritium; C023505:2',5'-oligoadenylate; D015088:2',5'-Oligoadenylate Synthetase; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease