Incorporation of lysine into Y base of phenylalanine tRNA in Vero cells.
Vero cells, a line derived from African green monkey kidney, contains a hypermodified base, called Y, adjacent to the 3' end of the anticodon of tRNAPhe. Two types of evidence are presented suggesting that lysine is involved in biosynthesis of Y base in these cells. First, when Vero cells are starved for lysine, a new, early-eluting species of tRNAPhe which lacks the fully modified Y base can be detected by reversed phase chromatography (RPC-5). After addition of lysine to the medium, this new species disappears. Second, when these cells are grown in low-lysine medium and then exposed to [3H]lysine, radioactivity from the lysine comigrates with tRNAPhe. The Y base can be selectively excised from tRNAPhe by incubation at pH 2.9, and extracted into ethyl acetate. Thin-layer chromatography of acid-excised material from these cells reveals that lysine-derived radioactivity comigrates with genuine Y base from calf liver tRNAPhe and the acid-excised tRNA no longer contains radioactivity. These results are consistent with the model that lysine is a structural precursor of Y base in tRNAPhe of Vero cells.
D000818:Animals; D000926:Anticodon; D002460:Cell Line; D006147:Guanine / Q000031:analogs & derivatives; D000882:Haplorhini; D007668:Kidney; D008239:Lysine / Q000378:metabolism*; D010649:Phenylalanine; D011688:Purinones / Q000096:biosynthesis*; D012343:RNA, Transfer / Q000096:biosynthesis*
D000926:Anticodon; D011688:Purinones; C012099:wye; D010649:Phenylalanine; D006147:Guanine; D012343:RNA, Transfer; D008239:Lysine