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Russian Journal of Genetics

, Volume 50, Issue 3, pp 319–322 | Cite as

Bacillus subtilis ypaA gene regulation mechanism by FMN riboswitch

  • S. A. Sklyarova
  • A. S. Mironov
Short Communications

Abstract

We studied the regulation of the Bacillus subtilis ypaA gene-encoding riboflavin-transporter by FMN riboswitch. Using translational fusions of the leader region of wild-type ypaA gene with the lacZ-reporter gene in the leader region we showed that in vivo ypaA gene expression decreased more than 10-fold in the presence of endogenous FMN. Introduction of two nucleotide substitutions providing stabilization of the sequester hairpins results in almost complete repression of reporter gene expression. Using toeprint assay in vitro it has been shown that FMN presence inhibits the formation of the 30S initiation complexint the ypaA gene leader mRNA. Our results support the model of ypaA gene regulation whereby FMN binding with the ypaA gene leader sequence results in translation suppression through the sequestering of the SD-sequence.

Keywords

Leader Region Translational Fusion Galactosidase Activity Flavin Mononucleotide Riboflavin Biosynthesis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Kreneva, R.A., Gel’fand, M.S., Mironov, A.A., et al., Inactivation of the ypaA gene in Bacillus subtilis: analysis of the resulting phenotypic expression, Russ. J. Genet., 2000, vol. 36, no. 8, pp. 972–974.Google Scholar
  2. 2.
    Vogl, C., Grill, S., Schilling, O., et al., Characterization of riboflavin (vitamin B2) transport proteins from Bacillus subtilis and Corynebacterium glutamicum, J. Bacteriol., 2007, vol. 189, no. 20, pp. 7367–7375.PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Ott, E., Stolz, J., Lehmann, M., et al., The RFN riboswitch of Bacillus subtilis is a target for the antibiotic roseoflavin produced by Streptomyces davawensis, RNA Biol., 2009, vol. 6, no. 3, pp. 276–280.PubMedCrossRefGoogle Scholar
  4. 4.
    Gelfand, M.S., 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, no. 11, pp. 439–442.PubMedCrossRefGoogle Scholar
  5. 5.
    Vitreschak, A.H., Rodionov, D.A., Mironov, A.A., and Gelfand, M.S., Regulation of riboflavin biosynthesis and transport genes in bacteria by transcriptional and translational attenuation, Nucleic Acids Res., 2002, vol. 30, pp. 3141–3151.PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    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, no. 5, pp. 747–756.PubMedCrossRefGoogle Scholar
  7. 7.
    Winkler, W.C., Cohen-Chalamish, S., and Breaker, R.R., An mRNA structure that controls gene expression by binding FMN, Proc. Natl. Acad. Sci. USA, 2002, vol. 99, no. 25, pp. 15908–15913.PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Bresler, S.E., Glazunov, E.A., Chernik, T.P., et al., Riboflavin biosynthesis operon of Bacillus subtilis: 5. Flavin mononucleotide and flavin adenine dinucleotide as effectors in the operon of riboflavin biosynthesis, Genetika (Moscow), 1973, vol. 9, no. 3, pp. 84–92.Google Scholar
  9. 9.
    Miller, J.H., A Short Course in Bacterial Genetics, New York: Cold Spring Harbor Lab., 1992.Google Scholar
  10. 10.
    Hartz, D., McPheeters, D.S., Traut, R., et al., Extension inhibition analysis of translation initiation complexes, Methods Enzymol., 1988, vol. 164, pp. 419–425.PubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Inc. 2014

Authors and Affiliations

  1. 1.State Research Institute of Genetics and Selection of Industrial MicroorganismsMoscowRussia

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