Skip to main content
Log in

Functional characterization of the cutI gene for the transcription of carbon monoxide dehydrogenase genes in Mycobacterium sp. strain JC1 DSM 3803

  • Microbial Genetics, Genomics and Molecular Biology
  • Published:
Journal of Microbiology Aims and scope Submit manuscript

Abstract

Carbon monoxide dehydrogenase (CO-DH) in Mycobacterium sp. strain JC1 is a key enzyme for the carboxydotrophic growth, when carbon monoxide (CO) is supplied as a sole source of carbon and energy. This enzyme is also known to act as nitric oxide dehydrogenase (NO-DH) for the detoxification of NO. Several accessory genes such as cutD, cutE, cutF, cutG, cutH, and cutI, are clustered together with two copies of the CO-DH structural genes (cutB1C1A1 and cutB2C2A2) in Mycobacterium sp. strain JC1 and are well conserved in carboxydotrophic mycobacteria. Transcription of the CO-DH structural and accessory genes was demonstrated to be increased significantly by acidified sodium nitrate as a source of NO. A cutI deletion (ΔcutI) mutant of Mycobacterium sp. strain JC1 was generated to identity the function of CutI. Lithoautotrophic growth of the ΔcutI mutant was severely affected in mineral medium supplemented with CO, while the mutant grew normally with glucose. Western blotting, CO-DH activity staining, and CO-DH-specific enzyme assay revealed a significant decrease in the cellular level of CO-DH in the ΔcutI mutant. Northern blot analysis and promoter assay showed that expression of the cutB1 and cutB2 genes was significantly reduced at the transcriptional level in the ΔcutI mutant, compared to that of the wildtype strain. The ΔcutI mutant was much more susceptible to NO than was the wild type.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bryk, R., Griffin, P., and Nathan, C. 2000. Peroxynitrite reductase activity of bacterial peroxiredoxins. Nature 407, 211–215.

    Article  CAS  PubMed  Google Scholar 

  • Cho, J.W., Yim, H.S., and Kim, Y.M. 1985. Acinetobacter isolate growing with carbon monoxide. Korean J. Microbiol. 23, 1–8.

    CAS  Google Scholar 

  • Howard, N.S., Gomez, J.E., Ko, C., and Bishai, W.R. 1995. Color selection with a hygromycin-resistance-based Escherichia coli-mycobacterial shuttle vector. Gene 166, 181–182.

    Article  CAS  PubMed  Google Scholar 

  • Hu, Y.M., Butcher, P.D., Mangan, J.A., Rajandream, M.A., and Coates, A.R.M. 1999. Regulation of hmp gene transcription in Mycobacterium tuberculosis: effects of oxygen limitation and nitrosative and oxidative stress. J. Bacteriol. 181, 3486–3493.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kerby, R.L., Youn, H., and Roberts, G.P. 2008. RcoM: a new singlecomponent transcriptional regulator of CO metabolism in bacteria. J. Bacteriol. 190, 3336–3343.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim, Y.M. and Hegeman, G.D. 1981. Purification and some properties of carbon monoxide dehydrogenase from Pseudomonas carboxydohydrogena. J. Bacteriol. 148, 904–911.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kim, Y.M. and Hegeman, G.D. 1983. Oxidation of carbon monoxide by bacteria. Int. Rev. Cytol. 81, 1–32.

    Article  CAS  PubMed  Google Scholar 

  • Kim, Y.M. and Park, S.W. 2012. Microbiology and genetics of CO utilization in mycobacteria. Antonie van Leeuwenhoek 101, 685–700.

    Article  CAS  PubMed  Google Scholar 

  • King, G.M. and Weber, C.F. 2007. Distribution, diversity and ecology of aerobic CO-oxidizing bacteria. Nat. Rev. Microbiol. 5, 107–118.

    Article  CAS  PubMed  Google Scholar 

  • Kraut, M., Hugendieck, I., Herwig, S., and Meyer, O. 1989. Homology and distribution of CO dehydrogenase structural genes in carboxydotrophic bacteria. Arch. Microbiol. 152, 335–341.

    Article  CAS  PubMed  Google Scholar 

  • Lee, J.H., Park, D.O., Park, S.W., Hwang, E.H., Oh, J.I., and Kim, Y.M. 2009. Expression and regulation of ribulose 1,5-bisphosphate carboxylase/ oxygenase genes in Mycobacterium sp. strain JC1 DSM 3803. J. Microbiol. 47, 297–307.

    Article  PubMed  Google Scholar 

  • Meyer, O. and Schlegel, H.G. 1983. Biology of aerobic carbon monoxide- oxidizing bacteria. Annu. Rev. Microbiol. 37, 277–310.

    Article  CAS  PubMed  Google Scholar 

  • Miller, J.H. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.

    Google Scholar 

  • Oh, J.I., Park, S.J., Shin, S.J., Ko, I.J., Han, S.J., Park, S.W., Song, T., and Kim, Y.M. 2010. Identification of trans- and cis-control elements involved in regulation of the carbon monoxide dehydrogenase genes in Mycobacterium sp. strain JC1 DSM 3803. J. Bacteriol. 192, 3925–3933.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ouellett, H., Ouellett, Y., Richard, C., Labarre, M., Wittenberg, B., Wittenberg, J., and Guertin, M. 2002. Truncated hemoglobin HbN protects Mycobacterium bovis from nitric oxide. Proc. Natl. Acad. Sci. USA 99, 5902–5907.

    Article  Google Scholar 

  • Parish, T. and Stoker, N.G. 1998. Electroporation of mycobacteria. In Parish, T. and Stoker, N.G. (eds.), Methods in Molecular Biology, pp. 129–144. Mycobacteria Protocols Humana Press, Totowa, New Jersey, USA.

    Google Scholar 

  • Park, S.W., Hwang, E.H., Jang, H.S., Lee, J.H., Kang, B.S., Oh, J.I., and Kim, Y.M. 2009. Presence of duplicate genes encoding a phylogenetically new subgroup of form I ribulose 1,5-bisphosphate carboxylase/oxygenase in Mycobacterium sp. strain JC1 DSM 3803. Res. Microbiol. 160, 159–165.

    Article  CAS  PubMed  Google Scholar 

  • Park, S.W., Hwang, E.H., Park, H., Kim, J.A., Heo, J., Lee, K.H., Song, T., Kim, E., Ro, Y.T., Kim, S.W., et al. 2003. Growth of mycobacteria on carbon monoxide and methanol. J. Bacteriol. 185, 142–147.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Park, S.W., Song, T., Kim, S.Y., Kim, E., Oh, J.I., Eom, C.Y., and Kim, Y.M. 2007. Carbon monoxide dehydrogenase in mycobacteria possesses a nitric oxide dehydrogenase activity. Biochem. Biophys. Res. Commun. 362, 449–453.

    Article  CAS  PubMed  Google Scholar 

  • Pelzmann, A., Ferner, M., Gnida, M., Meyer-Klaucke, W., Maisel, T., and Meyer, O. 2009. The CoxD protein of Oligotropha carboxidovorans is a predicted AAA plus ATPase chaperone involved in the biogenesis of the CO dehydrogenase [CuSMoO2] cluster. J. Biol. Chem. 284, 9578–9586.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pelzmann, A.M., Mickoleit, F., and Meyer, O. 2014. Insights into the posttranslational assembly of the Mo-, S- and Cu-containing cluster in the active site of CO dehydrogenase of Oligotropha carboxidovorans. J. Biol. Inorg. Chem. 19, 1399–1414.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roberts, G.P., Youn, H., and Kerby, R.L. 2004. CO-sensing mechanisms. Microbiol. Mol. Biol. Rev. 68, 453–473.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sambrook, J. and Russell, D.W. 2001. Molecular cloning: a laboratory manual, 3rd. Ed. Cold Spring Harbor Laboratory, NY, USA.

    Google Scholar 

  • Santiago, B., Schubel, U., Egelseer, C., and Meyer, O. 1999. Sequence analysis, characterization and CO-specific transcription of the cox gene cluster on the megaplasmid pHCG3 of Oligotropha carboxidovorans. Gene 236, 115–124.

    Article  CAS  PubMed  Google Scholar 

  • Shelver, D., Kerby, R.L., He, Y.P., and Roberts, G.P. 1997. CooA, a CO-sensing transcription factor from Rhodospirillum rubrum, is a CO-binding heme protein. Proc. Natl. Acad. Sci. USA 94, 11216–11220.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sherman, D.R., Voskuil, M., Schnappinger, D., Liao, R.L., Harrell, M.I., and Schoolnik, G.K. 2001. Regulation of the Mycobacterium tuberculosis hypoxic response gene encoding alpha-crystallin. Proc. Natl. Acad. Sci. USA 98, 7534–7539.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Song, T., Lee, H., Park, Y.H., Kim, E., Ro, Y.T., Kim, S.W., and Kim, Y.M. 2002. Reclassification of a carboxydobacterium, Acinetobacter sp. strain JC1 DSM3803, as Mycobacterium sp. strain JC1 DSM 3803. J. Microbiol. 40, 237–240.

    CAS  Google Scholar 

  • Song, T., Park, S.W., Park, S.J., Kim, J.H., Yu, J.Y., Oh, J.I., and Kim, Y.M. 2010. Cloning and expression analysis of the duplicated genes for carbon monoxide dehydrogenase of Mycobacterium sp. strain JC1 DSM 3803. Microbiology 156, 999–1008.

    Article  CAS  PubMed  Google Scholar 

  • Wilson, K. 1989. Preparation of genomic DNA from bacteria. In Ausubel, F.A., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., and Struhl, K. (eds.), Current Protocols in Molecular Biology, pp. 241–245. Greene Publishing Associates and Wiley-Interscience, NY, USA.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jeong-Il Oh.

Additional information

These authors contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lee, J.H., Park, S.W., Kim, Y.M. et al. Functional characterization of the cutI gene for the transcription of carbon monoxide dehydrogenase genes in Mycobacterium sp. strain JC1 DSM 3803. J Microbiol. 55, 31–36 (2017). https://doi.org/10.1007/s12275-017-6572-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12275-017-6572-7

Keywords

Navigation