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

Abstract

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.