Adenylation of Small RNAs in Human Cells DEVELOPMENT OF A CELL-FREE SYSTEM FOR ACCURATE ADENYLATION ON THE 39-END OF HUMAN

The 3*-end sequences of several human small RNAs were determined, and the results show that a fraction of human cytoplasmic 7SL, ribosomal 5S, and nuclear U2, U6, and 7SK small RNAs contain a post-transcriptionally added adenylic acid residue on their 3*-ends. Incubation of HeLa cell extract in vitro in the presence of [a-P]ATP resulted in labeling of several small RNAs including ribosomal 5S and cytoplasmic 7SL as well as U2 and U6 small nuclear RNAs. Analysis of 7SL RNA labeled in this in vitro adenylation system showed that a single adenylic acid residue is added to the 3*-end. These results show that the adenylation observed in the in vitro system reflects the post-transcriptional adenylation occurring in vivo.