In Vivo Aminoacylation of Transfer Ribonucleic Acid in Bacillus subtilis and Evidence for Differential Utilization of Lysine-Isoaccepting Transfer Ribonucleic Acid Species

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Abstract
The presence or absence of certain amino acids had different effects on the ability of B. subtilis to sporulate, and the intracellular pool size of amino acids varies during sporulation. To determine if these variations exert a regulatory effect through aminoacylation of tRNA, the levels of aminoacylation in vivo in the log or stationary phase of growth were studied. Periodate oxidation method and amino acid analyzer were used to evaluate in vivo aminoacylation. In general the level of aminoacylation of tRNA remains constant through stage III of sporulation, although there were detectable variations for specific amino acid groups. Periodate oxidation damaged certain tRNA; the results obtained by such a method should thus be intepreted with caution. Since the damage can affect certain isoaccepting species specifically, the periodate oxidation method cannot be used to establish which isoaccepting species are acylated in vivo. A preferential use of lysyl-tRNA by polyribosomes at different growth stages was demonstrated. Control mechanisms operating during the early stages of sporulation, therefore, do not affect the overall level of aminoacylation. However, there is an effect on the levels of aminoacylation of specific amino acids and on which isoaccepting species are utilized by the polyribosome system.