Analysis of Isoaccepting Transfer Ribonucleic Acid Species of Bacillus subtilis : Chromatographic Differences Between Transfer Ribonucleic Acids from Spores and Cells in Exponential Growth

Abstract

Differences between the transfer ribonucleic acid (tRNA) of spores and exponentially growing cells of Bacillus subtilis 168 were compared by co-chromatography on reversed-phase column RPC-5. This system gave excellent resolution of isoaccepting species in 1 to 2 hr using a 200-ml gradient. Two methods were used to extract spore tRNAs, a procedure using a Braun homogenizer and a pretreatment with dithiothreitol followed by lysis with lysozyme. Where changes were observed, column elution profiles of spore tRNAs were independent of the extraction method used. Three kinds of changes between the profiles of vegetative cell tRNA and spore tRNA were observed: (i) no change; phe-, val-, ala-, asp-, ileu-, pro-, met-, fmet-, and his-tRNAs, (ii) a change in the ratio of existing peaks; gly-, tyr-, leu-, ser-, thr-, aspn-, and arg-tRNAs, and (iii) the appearance or disappearance of unique peaks; lys-, glu-, and trp-tRNAs.

Analysis of Isoaccepting Transfer Ribonucleic Acid Species of Bacillus subtilis: Changes in Chromatography of Transfer Ribonucleic Acids Associated with Stage of Development

“…The changes in the profiles of the asporogenous mutant resemble the changes found in the sporeforming strain for tryptophanyl-, leucyl-, and tyrosyl-tRNAs as well as the previously reported changes in threonyl-, glycyl-, seryl-, and arginyl-tRNAs (4). The results verify that these alterations are not artifacts of the method used to extract spores.…”

“…This paper presents a study of changes in the isoaccepting species of transfer ribonucleic acids (tRNAs) for tyrosine, leucine, lysine, and tryptophan as a function of the stage of development. These amino acid acceptor groups had previously been studied by using tRNAs isolated from cells in exponential growth and from spores, and all four groups had shown differences in chromatographic pattems (4). The present study reveals that the alterations in the isoaccepting tRNAs for tyrosine, leucine, lysine, and tryptophan do not all occur at the same stage of growth and may, therefore, be reflections of different developmental processes.…”

“…Chromatographic conditions. The same chromatographic conditions on reversed-phase column, tRNA CHANGES AS A FUNCTION OF DEVELOPMENT RPC-5, were used as for the previous study of tRNAs from exponentially growing cells and spores (4), except that 20 mM instead of 2 mM 2-mercaptoethanol was used in the elution buffers. The composition of the aminoacylation reactions was the same as reported (5) except that 1-ml reactions were used and the concentration of 2-mercaptoethanol was raised to 10 mM.…”

Nucleotide sequence of a lysine tRNA from Bacillus subtilis

“…BaciZlus subtiZis changes in the percentage of isoaccepting species of tRNAs in different growth stages or in different growth media [1][2][3][4][5][6][7][8][9][10][11]. Several authors have shown that there are two major isoaccepting species of lysine tRNAs in B. subtiZis and that percentage of these species varies in vegetative cells and spores [2,[7][8][9][10][11]. Using lysyl-tRNA-DNA hybridization technique, Chuang et aZ.…”

Properties and Developmental Roles of the Lysyl- and Tryptophanyl-Transfer Ribonucleic Acid Synthetases of Bacillus subtilis : Common Genetic Origin of the Corresponding Spore and Vegetative Enzymes

“…(ii) L-Lysine and its immediate precursor, meso-diaminopimelic acid are major constituents of the spore coat and spore cortex (34,37). (iii) The tRNAlYs iso-accepting species undergo both quantitative and qualitative changes at specific times during sporogenesis (18,43,44). Although some asporogenic mutants do not show these changes, others are blocked in these events (19,44).…”