1887

Abstract

The single ribosomal RNA () operons of slow-growing mycobacteria comprise the genes for 16S, 23S and 5S rRNA, in that order. PCR methodology was used to amplify parts of the operons, namely the spacer-1 region separating the 16S rRNA and 23S rRNA genes and the spacer-2 region separating the 23S rRNA and 5S rRNA genes of . The amplified DNA was sequenced. The spacer-2 region, the 5S rRNA gene, the trailer region and the downstream region of the operon of were cloned and sequenced. These data, together with those obtained previously for , were used to identify putative antitermination signals and RNase III processing sites within the spacer-1 region. Notable features include two adjacent potential Box B elements and a Box A element. The latter is located within a sequence of 46 nucleotides which is very highly conserved among the slow-growers which were examined. The conserved sequence has the capacity to interact through base-pairing with part of the spacer-2 region. Secondary structures for mycobacterial precursor 23S rRNA and for precursor 5S rRNA were devised, based on sequence homologies and homologous nucleotide substitutions. All the slow-growers, including , conform to the same scheme of secondary structure. A putative motif for the intrinsic termination of transcription was identified approximately 33 bp downstream from the 3'-end of the 5S rRNA gene. The spacer-1 and spacer-2 sequences may prove a useful supplement to 16S rRNA sequences in establishing phylogenetic relationships between very closely related species.

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1994-07-01
2024-05-03
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References

  1. Albrechtsen B., Squires C L., Squires C. 1990; Antitermination of characterized transcriptional terminators by the Escherichia coli rrn G leader region. J Mol Biol 213:123–134
    [Google Scholar]
  2. Apirion D., Miczak A. 1993; RNA Processing in prokaryotic cells. Bio Essays 15:113–120
    [Google Scholar]
  3. Baylis H.A., Bibb M. J. 1988; Transcriptional analysis of the 16S rRN A gene of the rrn D gene set of Streptomyces coelicolor A3(2). Mol Microbiol 2:569–579
    [Google Scholar]
  4. Berg K. L., Squires C., Squires C. L. 1989; Ribosomal RN A operon anti-termination: function of leader and spacer region Box B Box A sequences and their conservation in diverse microorganisms. J Mol Biol 209:345–358
    [Google Scholar]
  5. Bercovier H., Kafri O., Sela S. 1986; Mycobacteria possess a surprisingly small number of ribosomal RNA genes in relation to the size of their genome. Biochem Biopbys Res Commun 136:1136–1141
    [Google Scholar]
  6. Bruckner D.A., Colonna P. 1993; Nomenclature for aerobic and facultative bacteria. Clin Infect Dis 16:598–605
    [Google Scholar]
  7. Dams E., Yamada T., De Baene R., Huysmans E., Vandenberghe A., De Wachter R. 1987; Structure of 5S rRNA in actinomycetes and relatives and evolution of eubacteria. J Mol Evol 25:255–260
    [Google Scholar]
  8. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395
    [Google Scholar]
  9. Dubos R.J., Davis B. D. 1946; Factors affecting the growth of tubercle bacilli in liquid media. J Exp Med 83:409–423
    [Google Scholar]
  10. Ehresmann C. P., Carpousis A. J., Krisch H. M. 1992; Specificity of Escherichia coli endoribonuclease RNase E: in vivo and in vitro analysis of mutants in a bacteriophage T4 mRN A processing site. Genes and Dev 6:149–159
    [Google Scholar]
  11. Fotheringham R., Wilson K. 1993; Sequence-based differentiation of strains in the Mycobacterium avium complex. J Bacteriol 175: 2818; 2825:
    [Google Scholar]
  12. Ghora B.K., Apirion D. 1978; Structural analysis and in vitro processing to P5 rRNA of a 9S RNA molecule isolated from an rrn mutant of E. coli . Cell 15:1055–1066
    [Google Scholar]
  13. Gutell R. R., Schnare M. N., Gray M. W. 1990; A compilation of large subunit (23S-like) ribosomal RNA sequences presented in a secondary structure format. Nucleic Acids Res 18:2319–2331
    [Google Scholar]
  14. Iwami M., Muto A., Yamao F., Osawa S. 1984; Nucleotide sequence of the rrn B 16S ribosomal RNA gene from Mycoplasma capricolum . Mol and Gen Genet 196:317–322
    [Google Scholar]
  15. Ji Y.-E., Colton M. J., Cox R. A. 1994; Nucleotide sequence and secondary structures of precursor 16S rRNA of slow-growing mycobacteria. Microbiology 140:123–132
    [Google Scholar]
  16. Kempsell K.E., Ji Y.-E., Estrada G, I. C. E., Colton M. J., Cox R. A. 1992; The nucleotide sequence of the promoter, 16S rRNA and spacer region of the ribosomal RNA operon of Mycobacterium tuberculosis and comparison with Mycobacterium leprae precursor rRNA. J Gen Microbiol 138:1717–1727
    [Google Scholar]
  17. Liesack W., Sela S., Bercovier H., Pitulle C., Stackebrandt E. 1991; G unplete nucleotide sequence ot the Mycobacterium leprae 23S and 5S rRNA genes plus flanking regions and their potential in designing diagnostic oligonucleotide probes. FEBS Lett 281:114–118
    [Google Scholar]
  18. Lindahl L., Zengel J. M. 1979; Operon-specific regulation of ribosomal protein synthesis in Escherichia coli . Proc Natl Acad Sci U.V. 176:6542–6546
    [Google Scholar]
  19. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Meissner G., Schroder K.-H. 1975; Relationship between Mycobacterium simiae and Mycobacterium habana . Am Rer Respir Dis 111:196–200
    [Google Scholar]
  21. Ninio J. 1979; Prediction of pairing schemes in RNA molecules loop contributions and energy of wobble and non-wobble base-pairs. Biochemistry 61:1133–1150
    [Google Scholar]
  22. Nodwell J.R., Greenblatt J. 1993; Recognition of Box A antiterminator RNA bv the E. coli antitermination factors Nus B and ribosomal protein S10. Cell 72:261–268
    [Google Scholar]
  23. Ogasawara N., Moriya S., Yoshikawa H. 1983; Structure and organization of rRNA operons in the region of the replication origin ot the Bacillus subti/is chromosome. Nucleic Acids Res 11:6301–6318
    [Google Scholar]
  24. Ohkubo S., Lwasaki H., Hori H., Osowa S. 1986; Evolutionary relationship of denitrifying bacteria as deduced from 5S rRNA sequences. J Biochem 100:1261–1267
    [Google Scholar]
  25. Pernodet J.-L., Boocard F., Alegre M.-T., Gagnat J., Guerineau M. 1989; Organization and nucleotide sequence analysis of a ribosomal RNA cluster from Streptomyces ambofaciens. Gene 79:33–46
    [Google Scholar]
  26. Portaels F. 1980; Unclassified mycobacterial strains susceptible to dapsone isolated from the environment in central Africa. Int J Lepr Other Mycobact Dis 48:330
    [Google Scholar]
  27. Richardson J.P. 1993; Transcription termination. Crit Rer Biochem Mol Biol 28:130
    [Google Scholar]
  28. Roberts J.W. 1993; RNA and protein elements of E. coli and λ transcription antitermination complexes. Cell 72:653–655
    [Google Scholar]
  29. Sela S., Clark-Curtiss J.E. 1991; Cloning and characterization of the Mycobacterium leprae putative ribosomal RNA promoter in Escherichia coli . Gene 98:123–127
    [Google Scholar]
  30. Shepard C.C. 1960; T he experimental disease that follows the injection of human leprosy bacilli into the footpads of mice. J Exp Med 112:445–454
    [Google Scholar]
  31. Singh N. B., Lowe A. C. R. E., Rees R. J. W., Colston M. J. 1989; Vaccination of mice against Mycobacterium leprae infection. Infect Immun 57:653–655
    [Google Scholar]
  32. Suzuki Y., Nagata A., Ono Y., Yamada I. 1988; Complete nucleotide sequence of the 16S rRNA gene of Mycobacterium bovis BCG. J Bacteriol 170:1631 –1636
    [Google Scholar]
  33. Taschke C., Herrmann R. 1986; Analysis of transcription and processing signals of the 16S 23S rRNA operon of Mycoplasma byopneumoniae. Mol and Gen Genet 205:434–441
    [Google Scholar]
  34. Valdivia Alvarez F., Suarez Mendez R., Echemendia Font M. 1971; Mycobacterium habana: probable especie dentro de las micobacterias no clasirtcadas. Bot Hig Epidemiol 9:65–73
    [Google Scholar]
  35. Vogelstein B., Gillespie D. 1979; Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci USA 76:615–619
    [Google Scholar]
  36. Von Hippel P.H., Yager T. D. 1991; Transcript elongation and termination are competitive kinetic processes. Proc Natl Acad Sci USA 88:2307–2311
    [Google Scholar]
  37. Walters J., Erdmann V. A. 1988; Compilation of 5S rRNA and 5S rRNA gene sequences. Nucleic Acids Res 18:2215–2236
    [Google Scholar]
  38. Winder F.G., Rooney S. A. 1970; I vffects of nitrogenous components of the medium on the carbohydrate and nucleic acid content of Mycobacterium tuberculosis BCG . J Gen Microbiol 63:29–39
    [Google Scholar]
  39. Zengel J. M., Muecki D., Lindahl L. 1980; Protein L4 of the E. coli ribosome regulates an eleven gene rprotein operon. Cell 21:523–535
    [Google Scholar]
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