No abstract

Recombinant DNA and classic genetic procedures were used to map a spectinomycin resistance mutation to a 121 base pair region of a 16S RNA gene and a macrolide-lincosamide-streptogramin type B resistance mutation to a 32 base pair region of a 23S RNA gene. DNA sequence analysis of these regions revealed that spectinomycin resistance results from a C/G to T/A transition at position 1192 of a 16S RNA gene. Resistance to macrolide, lincosamide and streptogramin type B antibiotics results from an A/T to T/A transversion at position 2058 of a 23S RNA gene. The alteration in 16S RNA is in a sequence that can participate in alternate base pairing arrangements that have been proposed to be involved in the translocation process. The alteration in 23S RNA identifies sequences important to peptidyl transfer.

Transfer RNA genes ("spacer tRNA genes") are present in the spacer region between 16S and 23S rRNA genes in Escherichia coli. We have analyzed spacer tRNA genes carried by seven rRNA operons with different chromosomal locations. Six of these were isolated on plasmids and one on a transducing phage. We found that, in addition to the two previously identified genes for tRNA2Glu and tRNAIIle, there is a spacer tRNA gene which codes for tRNAIBAla. Of the seven rRNA operons studied, three had both tRNAIBAla and tRNAIIle genes, and the remaining four had the tRNA2Glu gene in their spacers. In addition, genes for tRNAIAsp were found near the distal ends of two different rRNA operons.

No abstract

Insertions of the transposon Tn 10 have been obtained in an E. coli ribosomal RNA operon (rrnX). These insertions were used to determine whether present models of polarity are sufficient to explain transcriptional properties of operon which are transcribed but not translated. Most models of polarity suggest that transcription of mRNA requires simultaneous translation of nascent mRNA, or else premature termination of transcription will result. This model as stated does not encompass rrn operons. In this study three Tn 10 insertions into rrnX were characterized. All three Tn 10 insertions do not completely eliminate in vivo transcription of the most distal tRNA gene of rrnX. In ultraviolet-irradiated cells one Tn 10 insertion reduces synthesis of downstream portions of rrnX by at most 2--3 fold. Two other insertions may have more extensive polar effects, although the exact level of polarity is difficult to evaluate because these Tn 10 insertions may affect RNA maturation and stability.