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Geobacillus stearothermophilus V ubiE gene product is involved in the evolution of dimethyl telluride in Escherichia coli K-12 cultures amended with potassium tellurate but not with potassium tellurite

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Abstract

A 3.8-kb fragment of chromosomal DNA of Geobacillus stearothermophilus V cloned in pSP72 (p1VH) confers resistance to potassium tellurite (K2TeO3) and to potassium tellurate (K2TeO4) when the encoded genes are expressed in Escherichia coli K-12. The nt sequence of the cloned fragment predicts three ORFs of 780, 399, and 600 bp, whose encoded protein products exhibit about 80% similarity with the SUMT methyltransferase and the BtuR protein of Bacillus megaterium, and with the UbiE methyltransferase of Bacillus anthracis A2012, respectively. In addition, E. coli/p1VH cells evolved dimethyl telluride, which was released into the headspace gas above liquid cultures when amended with K2TeO3 or with K2TeO4. After 48 h of growth in the presence of these compounds, a protein of about 25 kDa was found at a significantly higher level when crude extracts were analyzed by SDS-PAGE. The N-terminal amino acid (aa) sequence of this protein, obtained by Edman degradation, matched the deduced aa sequence predicted by the G. stearothermophilus V ubiE gene. This gene was amplified by PCR, subcloned in pET21b, and transformed into E. coli JM109(DE3). Interestingly, DMTe evolution occurred when these modified cells were grown in K2TeO4 – but not in K2TeO3 – amended media. These results may be indicative that the two Te oxyanions could be detoxified in the cell by different metabolic pathways.

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Acknowledgements

This work was supported by a Robert A. Welch departmental grant (J.W.S., M.F.P., and T.G.C.) and by grant 1030234 from FONDECYT (Chile) to C.C.V. and by DICYT grants from Universidad de Santiago de Chile to C.P.S. and to C.C.V. M.A.A. was supported by a doctoral fellowship from MECESUP UCH 106 (Chile).

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Authors and Affiliations

  1. Laboratorio de Microbiología Molecular, Facultad de Química y Biología, Universidad de Santiago de Chile, Casilla 40, Correo 33 , Santiago, Chile

    Manuel A. Araya, Claudia P. Saavedra & Claudio C. Vásquez

  2. Department of Chemistry, Sam Houston State University, PO Box 2117, Huntsville, TX , 77341, USA

    Jerry W. Swearingen Jr., Mary F. Plishker & Thomas G. Chasteen

Authors
  1. Manuel A. Araya
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  2. Jerry W. Swearingen Jr.
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  3. Mary F. Plishker
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  4. Claudia P. Saavedra
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  5. Thomas G. Chasteen
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  6. Claudio C. Vásquez
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Correspondence to Claudio C. Vásquez.

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Araya, M.A., Swearingen, J.W., Plishker, M.F. et al. Geobacillus stearothermophilus V ubiE gene product is involved in the evolution of dimethyl telluride in Escherichia coli K-12 cultures amended with potassium tellurate but not with potassium tellurite. J Biol Inorg Chem 9, 609–615 (2004). https://doi.org/10.1007/s00775-004-0554-z

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  • DOI: https://doi.org/10.1007/s00775-004-0554-z

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