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Codon-anticodon interaction in tRNAPhe: II. A nuclear magnetic resonance study of the binding of the codon UUC
Authors:HAM Geerdes  JH Van Boom  CW Hilbers
Institution:1. Department of Biophysical Chemistry, University of Nijmegen Toernooiveld, 6525 ED Nijmegen The Netherlands;2. Gorlaeus Laboratoria, State University of Leiden P.O. Box 9502, 2300 AA Leiden The Netherlands
Abstract:The effect of binding of the codon UUC to yeast tRNAPhe was investigated by means of n.m.r.2 spectroscopy and analytical ultracentrifugation. Binding of UUC to the transfer RNA anticodon tends to promote the aggregation of tRNA molecules; this is manifest from a line broadening in the n.m.r. experiments as well as from an increase in s20,w the ultracentrifuge experiments. Such an aggregation of tRNA molecules was not observed upon addition of different oligonucleotides, as described in the accompanying paper. In addition to the general broadening observed in the n.m.r. spectra, specific resonances in the methyl proton spectrum as well as in the hydrogen-bonded proton spectrum are broadened or shifted upon binding of UUC.These results are explained on the basis of the premise that two different tRNA-UUC complexes can exist in solution. It is suggested that the binding of UUC tends to promote a disruption of the m7G46 · m22G22 base-pair and its neighbouring base-pairs.In studying the binding of U-U-U-U to yeast tRNAPhe no resonances of protons hydrogen-bonded between the oligonucleotide and the tRNA could be detected at low temperatures. This indicates, that at these temperatures the lifetime of the tRNA-U-U-U-U complex is substantially shorter than the lifetime of the other tRNA-oligonucleotide complexes studied in this and the accompanying paper under these conditions.
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