The thermophilic organisms were found to contain significantly less number of tRNAs compared with the other two groups, viz., mesophilic and psychrophilic (Fig. 1a). Such an observation is not unexpected as these thermophilic
organisms have to sustain a high temperature during their survival and are expected BI 6727 molecular weight to show ‘cost minimization’. The tRNAs that were significantly reduced had the anticodons of hydrophilic amino acids (Arg, Asn, Asp, Gln, Glu, Gly, Lys, Tyr, Val) while a few had anticodons for hydrophobic amino acids (Ile, Leu, Met, Phe) (Fig. 1b). The tRNAs that did not alter significantly were Ala-, Cys-, His-, Pro-, Ser-, Thr- and Trp-tRNA. Interestingly, none of the tRNAs showed any significant increase in number among the thermophilic organisms. The tRNA genes of thermophiles and hyperthermophiles exhibit a much higher GC content compared with mesophiles and psychrophiles. The GC content of tRNA genes shows a strong positive correlation with the OGT (r=0.85, P<0.0001). The higher GC content in the thermophilic and hyperthermophilic
group of organisms might be a strategy to facilitate intramolecular stabilization Selleck GSK126 of the RNA secondary structure at an elevated temperature. To examine this possibility, the secondary structures of tRNAs were determined through mfold at eight different temperatures, viz., 0, 10, 20, 30, 37, 50, 70 and 90 °C. Analysis of the entire data set revealed that tRNAs from psychrophilic organisms have a tendency to fold
with an unstable structure (more loops than stems) in a higher temperature range; the thermophiles and hyperthermophiles fold with a stable and similar structure in the entire range of temperature chosen, while mesophiles are between the above two groups. The results are shown in Fig. 2, which depicts representative secondary structure for Cys-tRNA and Phe-tRNA for three organisms: Methanopyrus kandleri AV19 (hyperthermophilic), Bacillus cereus E33L (mesophilic) and Psychrobacter Unoprostone arcticus 273-4 (psychrophilic). Thus, the thermophiles and hyperthermophiles have tRNA sequence preferences to adapt to the high temperature they thrive. Cluster analysis was applied to two sets of data from tRNA folding: the free energy of folding (dG) and the melting temperature (Tm). The folding was computed at eight different temperatures (0, 10, 20, 30, 37, 50, 70 and 90 °C) and the average dG/Tm for each organism was used in generating the clusters. The clustered images (Fig. 3) present large contiguous patches of color representing groups of organisms that share similar patterns of folding (represented by dG/Tm values) over a range of temperatures. To verify that the cluster is not an artifact of the clustering procedure, the procedure was repeated several times using the same data set, randomizing the order each time. However, the procedure yielded the same results every time.
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