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Structural similarities of tRNA's were compared using three sets of isoaccepting species that had previously been shown to undergo significant changes in chromatographic elution properties as a function of developmental stage inBacillus subtilis . Comparisons of the structures of the tRNA's were based on the composition of their modified nucleosides, comparisons of oligonucleotide elution profiles from RPC-5 columns, and two-dimensional electrophoretic fingerprint analysis of oligonucleotides. The tRNA's studied were tRNALys 1 and tRNALys 3 ; tRNATyr 1 and tRNATyr 2 ; and tRNATrp 1 and tRNATrp 2 . The results suggest that the difference among these pairs of isoaccepting species is a difference in the degree of post-transcriptional modifications of the anticodon loop region. The nucleosides involved wereN 6 -(Δ2 -isopentenyl)adenosine (i6 A), 2-methylthio-N 6 -(Δ2 -isopentenyl)adenosine (ms2 i6 A), and an unknown nucleoside K, which occurred in a position analogous toN -[9-(β-d -ribofuranosyl)purin-6-ylcarbamoyl]threonine. The amounts of i6 A and ms2 i6 A, determined using total tRNA from exponential-or stationary-phase cells, suggest that the thiomethylation of i6 A is a pleiotropic phenomenon affecting several tRNA species. As opposed to the situation inEscherichia coli tRNA, where ms2 i6 A constitutes about 90% of the total hydrophobic nucleosides at all growth stages,B. subtilis tRNA's have i6 A as the predominant hydrophobic nucleoside in exponential growth and ms2 i6 A as the predominant nucleoside in stationary phase. Thus, the enzyme system which forms i6 A and the enzyme system which thiomethylates i6 A are not coordinated during growth inB. subtilis as they are inE. coli . It is suggested that these changes in anticodon loop modifications inB. subtilis may be related to changes in the translational apparatus which occur during sporulation.

journal of bacteriology

Post-Transcriptional Modifications of the Anticodon Loop Region: Alterations in Isoaccepting Species of tRNA's During Development in <i>Bacillus subtilis</i>

Post-Transcriptional Modifications of the Anticodon Loop Region: Alterations in Isoaccepting Species of tRNA's During Development in Bacillus subtilis