A method for isolation of oligo(uridylic acid)-containing messenger ribonucleic acid from HeLa cells.
When cytoplasmic polyadenylated ribonucleic acid [poly(A+)RNA] from HeLa cells was treated with ribonuclease H (RNase H) and oligodeoxythymidylate [oligo(dT)] to remove its 3'-poly(A) tail, an increased binding to poly(A)-agarose was observed. The bound material, which comprised 4-6% of the initial RNA, contained 65-80% of the oligo(uridylic acid) [oligo(U)] sequences generated by RNase T1 digestion. Oligo(U) isolated from the bound fraction was shown to be 83% U and to have a U/G ratio of 33. In contrast, oligo(U) from the unbound material was 77% U and had a U/G ratio of 13, suggesting that it is shorter and less U rich than the oligo(U) in the bound fraction. On sucrose gradients, oligo(U+)RNA consistently sedimented with a larger s value than oligo(U-) RNA. The oligo(U) content of oligo(U+) RNA suggests one oligo(U) tract of 33 nucleotides per RNA molecule of 2000-3000 residues.
D001483:Base Sequence; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D006367:HeLa Cells / Q000032:analysis; D006801:Humans; D011061:Poly A / Q000302:isolation & purification; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger; D014500:Uracil Nucleotides
D012333:RNA, Messenger; D014500:Uracil Nucleotides; C032369:oligo(U+)RNA; D011061:Poly A; D012313:RNA