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The characterization of internal promoters in the Bacillus subtilis riboflavin biosynthesis operon

  • Genetics of Microorganisms
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Abstract

The transcription start sites of two internal promoters, the P2 and P3 promoters, in the Bacillus subtilis riboflavin biosynthesis operon were identified by primer extension. Putative −35 and −10 sequences that are recognized by the vegetative δ70 subunit of RNA polymerase have been found upstream of the P2 and P3 transcription start sites. The relative strengths of the P1, P2, and P3 promoters were determined by cloning these promoters into the pDG268 expression vector. It was shown that the transcriptional activity of the P3 promoter is approximately fivefold higher as compared with P1, the major promoter, whereas P2 promoter activity is lower by almost two orders of magnitude. Real-time PCR demonstrated that unlike the P1 promoter, P2- and P3-driven expression is not regulated by flavins.

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References

  1. Perkins, J.B. and Pero, J., Vitamin biosynthesis. Bacillus subtilis and Its Closest Relatives: From Genes to Cells, Sonenshein, A.L., Hoch, J.A., and Losick, R., Eds., Washington: American Society Microbiology Press, 2002, pp. 271–286.

    Google Scholar 

  2. Mironov, V.N., Kraev, A.S., Chikindas, M.L., et al., Functional Organization of the Riboflavin Biosynthesis Operon from Bacillus subtilis, Mol. Gen. Genet., 1994, vol. 242, pp. 201–208.

    Article  PubMed  CAS  Google Scholar 

  3. Mironov, A.S., Gusarov, I., Rafikov, R., et al., Sensing Small Molecules by Nascent RNA: A Mechanism to Control Transcription in Bacteria, Cell, 2002, vol. 111, pp. 747–756.

    Article  PubMed  CAS  Google Scholar 

  4. Kil, Y.V., Mironov, V.N., Gorishin, I.Y., et al., Riboflavin Operon in Bacillus subtilis: Unusual Symmetric Arrangement of the Regulatory Region, Mol. Gen. Genet., 1992, vol. 233, pp. 483–486.

    Article  PubMed  CAS  Google Scholar 

  5. Gelfand, M., Mironov A.A., Jomantas, J., et al., A Conserved RNA Structure Element Involved in the Regulation of Bacterial Riboflavin Synthesis Genes, Trends Genet., 1999, vol. 15, pp. 439–442.

    Article  PubMed  CAS  Google Scholar 

  6. Perkins, J.B., Sloma, A., Hermann, T., et al., Genetic Engineering of Bacillus subtilis for the Commercial Production of Riboflavin, Ind. Microbiol. Biotechnol., 1999, vol. 22, pp. 8–18.

    Article  CAS  Google Scholar 

  7. Stragier, P., Bonamy, C., and Karmazyn-Campelli, C., Processing of a Sporulation Sigma Factor in Bacillus subtilis: How Morphological Structure Could Control Gene Expression, Cell, 1988, vol. 52, pp. 697–704.

    Article  PubMed  CAS  Google Scholar 

  8. Vagner, V., Dervyn, E., and Ehrlich, S.D., A Vector for Systematic Gene Inactivation in Bacillus subtilis, Microbiology, 1998, vol. 144, pp. 3097–3104.

    Article  PubMed  CAS  Google Scholar 

  9. Miller, J., Experiments in Molecular Genetics, Cold Spring Harbor: Cold Spring Harbor Lab., 1972.

    Google Scholar 

  10. Saito, H. and Miura, K.I., Preparation of Transforming DNA by Phenol Treatment, Biochim. Biophys. Acta, 1963, vol. 42, pp. 619–629.

    Article  Google Scholar 

  11. Anagnostopoulos, C. and Spizizen, J., Requirements for Transformation in Bacillus subtilis, J. Bacteriol., 1961, vol. 81, pp. 741–746.

    PubMed  CAS  Google Scholar 

  12. Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Lab., 1989, 2ed.

    Google Scholar 

  13. Winestock, C.H., Aogaishi, T., and Plaut, G.W.E., The Substrate Specificity of Riboflavin Synthetase, J. Biol. Chem., 1963, vol. 238, pp. 2866–2874.

    PubMed  CAS  Google Scholar 

  14. Huntzinger, E., Possedko, M., Winter, F., et al., Probing RNA Structures with Enzymes and Chemicals in vitro and in vivo, Handbook of RNA Biochemistry, Hartmann, R.K., Bindereif, A., Schön, A., and Westhof, E., Eds., Weinheim: Wiley, 2008.

    Google Scholar 

  15. Jin, D.J., Slippage Synthesis at the galP2 Promoter of Escherichia coli and Its Regulation by UTP Concentration and cAMP-cAMP Receptor Protein, J. Biol. Chem., 1994, vol. 269, pp. 17221–17227.

    PubMed  CAS  Google Scholar 

  16. Ovcharova, I.V., Eremina, S.Yu., and Mironov, A.S., Determination of Functional Role of Nucleotide Composition in the Transcription Start Region of the Escherichia coli udp Gene, Russ. J. Genet., 2003, vol. 39, no. 3, pp. 256–263.

    Article  CAS  Google Scholar 

  17. Koh, S.A., Chung, W.H., Lee, J.H., and Roe, J.H., The Reversed SoxS-Binding Site Upstream of the ribA Promoter in Escherichia coli, Mol. Gen. Genet., 1999, vol. 261, pp. 374–380.

    Article  PubMed  CAS  Google Scholar 

  18. Berezovskii, V.M., Flavin Vitamins and Coenzymes, in Khimiya vitaminov (Vitamin Chemistry), Moscow, 1973, pp. 507–568.

  19. Serganov, A., Huang, L., and Patel, D.J., Coenzyme Recognition and Gene Regulation by a Flavin Mononucleotide Riboswitch, Nature, 2009, vol. 458, pp. 233–237.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Institute for Genetics and Selection of Industrial Microorganisms, Moscow, 113545, Russia

    S. A. Sklyarova & A. S. Mironov

  2. Konstantinov Petersburg Nuclear Physics Institute, Russian Academy of Sciences, Gatchina, Leningrad oblast, 188350, Russia

    R. A. Kreneva & D. A. Perumov

Authors
  1. S. A. Sklyarova
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  2. R. A. Kreneva
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  3. D. A. Perumov
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  4. A. S. Mironov
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Correspondence to A. S. Mironov.

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Original Russian Text © S.A. Sklyarova, R.A. Kreneva, D.A. Perumov, A.S. Mironov, 2012, published in Genetika, 2012, Vol. 48, No. 10, pp. 1133–1141.

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Sklyarova, S.A., Kreneva, R.A., Perumov, D.A. et al. The characterization of internal promoters in the Bacillus subtilis riboflavin biosynthesis operon. Russ J Genet 48, 967–974 (2012). https://doi.org/10.1134/S1022795412100109

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  • DOI: https://doi.org/10.1134/S1022795412100109

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