title: The binding of the 69- and 100-kD forms of 2',5'-oligoadenylate synthetase to different polynucleotides.
abstract: Three major forms of 2',5' oligoadenylate (2-5A) synthetase are induced in interferon (IFN)-treated human cells: 40-46, 69, and 100 kD. Here we studied the binding and activation of the 69- and 100-kD forms to single-stranded (ss) or double-stranded (ds) RNAs. The 69- and 100-kD form enzymes purified by immunoaffinity chromatography, were shown to be activated by synthetic dsRNAs poly(I).poly(C) or poly(A).poly(U) whereas ssRNAs poly(I), poly(C), poly(A), poly(U), and poly(G) had no effect. Both enzymes were also partially purified by binding to dsRNA or ssRNA-Sepharose. The synthetases bound to dsRNA Sepharose were partially activated but required the addition of soluble poly(I).poly(C) for maximal activity. The synthetases bound to ssRNA-Sepharose manifested no activity but became activated in the presence of soluble dsRNA, poly(I).poly(C). However, activation of such ssRNA-bound enzymes by dsRNA did not result in their dissociation from the ssRNA-Sepharose. These results indicate the presence of different polynucleotide binding sites on the 69- and 100-kD forms of 2-5A synthetase: a specific dsRNA binding site essential for activation and another polynucleotide binding site or sites which, although not specific, might be important for the optimal conformation of these proteins in cells.
authors.affiliation: Institut Pasteur, Unité de Virologie et Immunologie Cellulaire (URA CNRS 1157), Paris, France.
authors.forename: I
authors.initials: I
authors.lastname: Marié
authors.forename: J
authors.initials: J
authors.lastname: Svab
authors.forename: A G
authors.initials: AG
authors.lastname: Hovanessian
mesh_terms: D015088:2',5'-Oligoadenylate Synthetase / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D011131:Polyribonucleotides / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000378:metabolism
chemical_list: D011131:Polyribonucleotides; D012330:RNA, Double-Stranded; D012313:RNA; D015088:2',5'-Oligoadenylate Synthetase