Characterization of an rRNA Operon ( rrnB ) of Mycobacterium fortuitum and Other Mycobacterial Species: Implications for the Classification of Mycobacteria

Abstract
Mycobacteria are thought to have either one or two rRNA operons per genome. All mycobacteria investigated to date have an operon, designated rrnA , located downstream from the murA gene. We report that Mycobacteriun fortuitum has a second rrn operon, designated rrnB , which is located downstream from the tyrS gene; tyrS is very close to the 3" end of a gene ( 3-mag ) coding for 3-methylpurine-DNA-glycosylase. The second rrn operon of Mycobacterium smegmatis was shown to have a similar organization, namely, 5" 3-mag - tyrS - rrnB 3". The rrnB operon of M. fortuitum was found to have a single dedicated promoter. During exponential growth in a rich medium, the rrnB and rrnA operons were the major and minor contributors, respectively, to pre-rRNA synthesis. Genomic DNA was isolated from eight other fast-growing mycobacterial species. Samples were investigated by Southern blot analysis using probes for mur A, tyrS , and 16S rRNA sequences. The results revealed that both rrnA and rrnB operons were present in each species. The results form the basis for a proposed new scheme for the classification of mycobacteria. The approach, which is phylogenetic in concept, is based on particular properties of the rrn operons of a cell, namely, the number per genome and a feature of 16S rRNA gene sequences.