Nuclear magnetic resonance measurements of proton exchange were perfor
med on yeast tRNA(Phe) and yeast tRNA(Asp), at temperatures from 20 to
45 degrees C, in the presence of various levels of salt, phosphate, a
dded magnesium, and pH. The dynamical changes of the tRNA molecule wer
e interpreted, with the aid of firmly established assignments and the
use of the saturation recovery technique. In tRNA(Phe), the exchange r
ates in zero magnesium indicated early melting of the acceptor stem, t
ertiary structure, and D stem. However, in the presence of even low le
vels of magnesium the D stem remained intact up to high temperature, s
tabilized by a Mg2+ ion. A similar unfolding sequence was observed in
tRNA(Asp). The difference between these two tRNAs was the thermal beha
vior of the tertiary resonance U8-A14, In tRNA(Phe), this base pair sh
owed sharp rate increases between 32 and 39 degrees C. However, in tRN
A(Asp), it remained intact up to 36 degrees C and disappeared at 39 de
grees C, even if there was not important kinetic broadening. By measur
ing the temperature dependence of the exchange rates, we obtained an a
ctivation energy of 40-60 kcal/mol for all the imino protons of yeast
tRNAPhe in zero magnesium. The same activation energy was obtained for
tRNA(Phe) with equimolar concentration of magnesium. By investigation
of the dependence ofchange rates of these imino protons on solution c
onditions, we observed the transition from k(op) rate limiting in the
absence of magnesium to k(ex) rate limiting in the presence of magnesi
um.