Raman and 19F(1H) nuclear Overhauser evidence for a rigid solution conformation of Escherichia coli 5-fluorouracil 5S ribonucleic acid.

Escherichia coli cells grown on a medium containing 5-fluorouracil (FU) produce 5S RNA whose uracil residues are approximately 80% replaced by FU. The Raman spectra of native and FU-5S RNA are very similar, confirming similar solution conformations for the two species and a highly base-stacked structure in solution. The 254-MHz 19F NMR spectrum of FU-5S RNA shows that the 20-odd FU residues reside in at least ten distinct chemical environments, suggesting a highly ordered structure. Comparison of theoretical and experimental 19F(1H) nuclear Overhauser enhancements demonstrates definitively that virtually all the labeled uracils are bound to a rigid macromolecular frame, with a rotational correlation time of about 19 ns or longer. Since these uracils are widely distributed throughout the nucleotide primary sequence, it may be concluded that the entire FU-5S RNA solution structure is relatively rigid, in agreement with the most recently proposed "cloverleaf" secondary structural model for native prokaryotic 5S RNA.