Antibody nucleic acid complexes. Immunospecific retention of N6-methyladenosine-containing transfer ribonucleic acid.
Antibodies specific for N6-methyladenosine (m6A) were immobilized on Sepharose and the resulting immunoadsorbent was tested for its ability to retain those Escherichia coli tRNAs containing the antigenic hapten, i.e., m6A. Results obtained with [32P]PO4- and [methyl-3H]-methionine-labeled tRNAs indicated that approximately 3 to 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 (a) analysis of methyl-3H-labeled, methylated nucleosides, (b) two-dimensional gel electrophoresis, and (c) 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, namely, tRNAVal.
D000241:Adenosine / Q000031:analogs & derivatives* / Q000276:immunology; D000906:Antibodies; D002846:Chromatography, Affinity / Q000379:methods; D004926:Escherichia coli / Q000032:analysis; D006241:Haptens; D007163:Immunosorbent Techniques; D011233:Precipitin Tests; D012343:RNA, Transfer / Q000302:isolation & purification*; D012346:RNA, Transfer, Amino Acyl / Q000302:isolation & purification
D000906:Antibodies; D006241:Haptens; D012346:RNA, Transfer, Amino Acyl; D012343:RNA, Transfer; C010223:N-methyladenosine; D000241:Adenosine