tRNA modification profiles in obligate and moderate thermophilic bacilli.

Transfer RNAs (tRNAs) are the most ancient RNA molecules in the cell, modification pattern of which is linked to phylogeny. The aim of this study was to determine the tRNA modification profiles of obligate (Anoxybacillus, Geobacillus, Paragebacillus) and moderate (Bacillus, Brevibacillus, Ureibacillus, Paenibacillus) thermophilic aerobic bacilli strains to find out its linkage to phylogenetic variations between species. LC-MS was applied for the quantification of modified nucleosides using both natural and isotopically labeled standards. The presence of m²A and m⁷G modifications at high levels was determined in all species. Relatively high level of i⁶A and m⁵C modification was observed for Paenibacillus and Ureibacillus, respectively. The lowest level of Cm modification was found in Bacillus. The modification ms²i⁶A and m¹G were absent in Brevibacillus and Ureibacillus, respectively, while modifications Am and m²₂G were observed only for Ureibacillus. While both obligate and moderate thermophilic species contain Gm, m¹G and ms²i⁶A modifications, large quantities of them (especially Gm and ms²i⁶A modification) were detected in obligate thermophilic ones (Geobacillus, Paragebacillus and Anoxybacillus). The collective set of modified tRNA bases is genus-specific and linked to the phylogeny of bacilli. In addition, the dataset could be applied to distinguish obligate thermophilic bacilli from moderate ones.