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Nucleic Acids Research, 1979, Vol. 6, No. 6 2209-2216
© 1979

Articles

Incorporation of lysine into Y base of phenylalanine tRNA in Vero cells

Robert G. Pergolizzi, Dean L. Engelhardt and Dezider Grunberger

Cancer Center/Institute of Cancer Research, Departments of Biochemistry, Microbiology, and Division of Environmental Sciences, Columbia University College of Physicians and Surgeons New York, NY 10032, USA

Received March 12, 1979. 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 grnwn 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 tRRA 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.


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W. F. Waas, V. de Crecy-Lagard, and P. Schimmel
Discovery of a Gene Family Critical to Wyosine Base Formation in a Subset of Phenylalanine-specific Transfer RNAs
J. Biol. Chem., November 11, 2005; 280(45): 37616 - 37622.
[Abstract] [Full Text] [PDF]


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