Antibody-nucleic acid interactions. Antibodies to psoralen-modified RNA as probes of RNA structure.
Antisera elicited by immunization of rabbits with 4'-aminomethyl-trioxsalen (AMT)-modified poly(A,U) complexed with methylated bovine serum albumin was characterized in competition radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA). AMT-poly(A,U) was over 10,000-fold more reactive than unmodified poly(A,U) or AMT alone. The antiserum cross-reacted to varying extents with AMT-modified-RNA's and -DNA's. The presence of AMT-uridine usually assured strong reactivity. The amino group of AMT contributed to antibody binding to a small degree. Binding was not significantly affected by high ionic strength, suggesting that binding does not involve ion pair formation. Murine encephalomyocarditis virus replicative intermediates, as well as cellular RNA and DNA were modified by psoralen in intact cells, suggesting that EMCV RNA and cellular RNA's in intact cells possess detectable stretches of base pairs. The antibodies described here will be useful in studying the secondary and tertiary structure of RNA's in vitro and in intact cells.
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D003432:Cross-Linking Reagents; D004686:Encephalomyocarditis virus / Q000187:drug effects / Q000528:radiation effects; D004797:Enzyme-Linked Immunosorbent Assay; D007739:L Cells / Q000187:drug effects / Q000382:microbiology / Q000528:radiation effects; D009690:Nucleic Acid Conformation; D011063:Poly A-U / Q000276:immunology; D012313:RNA / Q000276:immunology*; D012330:RNA, Double-Stranded / Q000276:immunology; D012367:RNA, Viral / Q000187:drug effects / Q000276:immunology / Q000528:radiation effects; D011817:Rabbits; D011863:Radioimmunoassay; D012710:Serum Albumin, Bovine; D014307:Trioxsalen / Q000031:analogs & derivatives / Q000494:pharmacology; D014466:Ultraviolet Rays
D000906:Antibodies; D003432:Cross-Linking Reagents; D012330:RNA, Double-Stranded; D012367:RNA, Viral; C029180:methylated bovine serum albumin; D011063:Poly A-U; D012710:Serum Albumin, Bovine; C016507:hydroxymethyltrioxsalen; D012313:RNA; C018822:aminomethyltrioxsalen; D014307:Trioxsalen