Antibody nucleic acid complexes. Immunospecific retention of N6-methyladenosine-containing transfer ribonucleic acid

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Abstract
[Rabbit] antibodies specific for N6-methyladenosine (m6A) were immobilized on Sepharose and the resulting immunoadsorbent was tested for its ability to retain those Escherichia coli tRNA containing the antigenic hapten, i.e., m6A. Results obtained with [32P]PO4 and [methyl-3H]-methionine-labeled tRNAs indicated that approximately 3-5% of the radioactive RNA was retained by the immunoadsorbent. Under identical conditions, but in the presence of m6A (1 mg/ml), less than 0.2% of the radioactivity was retained. Subsequent characterization of the retained tRNA via analysis of methyl-3H-labeled, methylated nucleosides, 2-dimensional gel electrophoresis and analysis of the retention of [3H]aminoacyl-tRNA species led to the conclusion that the anti-m6A/Sepharose adsorbent quantitatively and exclusively retained a single tRNA species containing m6A (tRNAVal).