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We report here the finding of a highly significant inverse correlation of the uracil content of 16S rRNA and the optimum growth temperature (T(opt)) of cultured thermophilic and psychrophilic prokaryotes. This correlation was significantly different from the weaker correlations between the contents of other nucleotides and T(opt). Analysis of the 16S rRNA secondary structure regions revealed a fall in the A:U base-pair content in step with the increase in T(opt) that was much steeper than that of mismatched base-pairs, which are thermodynamically less stable. These findings indicate that the 16S rRNA sequences of thermophiles and psychrophiles are under a strong thermo-adaptive pressure, and that structure-function constraints play a crucial role in determining their 16S rRNA nucleotide composition. The derived relationship between uracil content and T(opt) was used to develop an algorithm to predict the T(opt) values of uncultured prokaryotes lacking cultured close relatives and belonging to the phyla predominantly containing thermophiles. This algorithm may be useful in guiding the design of cultivation conditions for hitherto uncultured microbes.

Uracil content of 16S rRNA of thermophilic and psychrophilic prokaryotes correlates inversely with their optimal growth temperatures