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Journal of Bacteriology, February 2002, p. 1078-1088, Vol. 184, No. 4
0021-9193/01/$04.00+0     DOI: 10.1128/jb.184.4.1078-1088.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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

M. C. Menendez,1 M. J. Garcia,1 M. C. Navarro,1 J. A. Gonzalez-y-Merchand,2 S. Rivera-Gutierrez,2 L. Garcia-Sanchez,1 and R. A. Cox3*

Departamento de Medicina Preventiva, Facultad de Medicina, Universidad Autonoma de Madrid, 28029-Madrid, Spain,1 Departamento de Microbiologia, Escuela Nacional de Ciencias Biologicas, IPN, Mexico D.F. 06400, Mexico,2 Division of Mycobacterial Research, National Institute for Medical Research, London NW7 1AA, United Kingdom3

Received 5 September 2001/ Accepted 15 November 2001

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 murA, 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.

* Corresponding author. Mailing address: Division of Mycobacterial Research, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, United Kingdom. Phone: 44 020 89 59 36 66. Fax: 44 020 89 06 44 77. E-mail: rcox{at}nimr.mrc.ac.uk.

Journal of Bacteriology, February 2002, p. 1078-1088, Vol. 184, No. 4
0021-9193/01/$04.00+0     DOI: 10.1128/jb.184.4.1078-1088.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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