Journal Mobile Options
Table of Contents
Vol. 12, No. 3-4, 2007
Issue release date: June 2007
J Mol Microbiol Biotechnol 2007;12:227–240

Contribution of Extracytoplasmic Function Sigma Factors to Transition Metal Homeostasis in Cupriavidus metallidurans Strain CH34

Grosse C. · Friedrich S. · Nies D.H.
Institut für Mikrobiologie, Martin-Luther-Universität Halle-Wittenberg, Halle, Deutschland

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Cupriavidus metallidurans strain CH34 is a highly metal-resistant bacterium that contains 11 sigma factors of the extracytoplasmic function (ECF) protein family, which can be subgrouped into the ECF:FecI 1, ECF:FecI 2, ECF:RpoE and ‘(ECF)’ clusters. To analyze the contribution of these 11 sigma factors to metal resistance, upregulation of the respective genes was measured by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR). As determined by RT-PCR, the ECF sigma factor genes were part of two- to tetra-cistronic operons, each containing genes for the sigma factor plus one or two antisigma factors. The three sigma factors RpoJ, RpoK and RpoI (ECF:FecI 1 cluster) were upregulated by Cu(II) and Ni(II), and under conditions of iron depletion. The other 8 ECF sigma factor genes were not induced by iron depletion. Strong upregulation of rpoJ and rpoK under iron depletion in a ΔrpoI mutant strain and close vicinity of rpoI to genes involved in iron siderophore metabolism marked RpoI as the primary ECF sigma factor for siderophore-mediated iron uptake. Genes for RpoO, RpoL and RpoM (ECF:FecI 2 cluster) were not upregulated by transition metal cations and influenced metal resistance only weakly. Concerning the two ‘(ECF)’ group proteins, rpoQ was strongly upregulated by Cu(II) and deletion of rpoR led to a small decrease in copper resistance. Of the three ECF:RpoE-encoding genes, rpoP was not transcribed under the conditions tested, cnrH was upregulated by Ni(II) and essential for nickel resistance as known before. RpoE was required for full metal resistance of C. metallidurans. None of these 11 sigma factors was essential for metal resistance mediated by the cobalt, zinc and cadmium resistance determinant czc, or for its expression. However, RpoI was essential for siderophore production in C. metallidurans, and, in addition to the known role of CnrH in nickel resistance, RpoE, RpoI, RpoJ, RpoK and maybe also RpoQ are required for the outstanding transition metal resistance of this bacterium.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Deretic V, Chandrasekharappa S, Gill JF, Chatterjee DK, Chakrabarty A: A set of cassettes and improved vectors for genetic and biochemical characterization of Pseudomonas genes. Gene 1987;57:61–72.
  2. Don RA, Pemberton JM: Porperties of six pesticide degradation plasmids isolated from Alcaligenes paradoxus and Alcaligenes eutrophus. J Bacteriol 1981;145:681–686.
  3. Egler M, Groβe C, Grass G, Nies DH: Role of ECF sigma factor RpoE in heavy metal resistance of Escherichia coli. J Bacteriol 2005;187:2297–2307.
  4. Goris J, De Vos P, Coenye T, Hoste B, Janssens D, Brim H, Diels L, Mergeay M, Kersters K, Vandamme P: Classification of metal-resistant bacteria from industrial biotopes as Ralstonia campinensis sp. nov., Ralstonia metallidurans sp. nov. and Ralstonia basilensis Steinleet al. 1998 emend. Int J Syst Evol Microbiol 2001;51:1773–1782.
  5. Grass G, Fricke B, Nies DH: Control of expression of a periplasmic nickel efflux pump by periplasmic nickel concentrations. Biometals 2005;18:437–448.
  6. Grass G, Groβe C, Nies DH: Regulation of the cnr cobalt/nickel resistance determinant from Ralstonia sp. CH34. J Bacteriol 2000;182:1390–1398.
  7. Grass G, Thakali K, Klebba PE, Thieme D, Müller A, Wildner GF, Rensing C: Linkage between catecholate siderophores and the multicopper oxidase CueO in Escherichia coli. J Bacteriol 2004;186:5826–5833.
  8. Groβe C, Anton A, Hoffmann T, Franke S, Schleuder G, Nies DH: Identification of a regulatory pathway that controls the heavy metal resistance system Czc via promoter czcNp in Ralstonia metallidurans. Arch Microbiol 2004;182:109–118.

    External Resources

  9. Groβe C, Grass G, Anton A, Franke S, Navarrete Santos A, Lawley B, Brown NL, Nies DH: Transcriptional organization of the czc heavy metal homoeostasis determinant from Alcaligenes eutrophus. J Bacteriol 1999;181:2385–2393.
  10. Gunesekere IC, Kahler CM, Ryan CS, Snyder LAS, Saunders NJ, Rood JI, Davies JK: Ecf, an alternative sigma factor from Neisseria gonorrhoeae, controls expression of msrAB, which encodes methionine sulfoxide reductase. J Bacteriol 2006;188:3463–3469.
  11. Helmann JD: The extracytoplasmic (ECF) sigma factors. Adv Microbiol Physiol 2002;46:47–110.
  12. Juhnke S, Peitzsch N, Hübener N, Groβe C, Nies DH: New genes involved in chromate resistance in Ralstonia metallidurans strain CH34. Arch Microbiol 2002;179:15–25.
  13. Liesegang H, Lemke K, Siddiqui RA, Schlegel HG: Characterization of the inducible nickel and cobalt resistance determinant cnr from pMOL28 of Alcaligenes eutrophus CH34. J Bacteriol 1993;175:767–778.
  14. Lonetto MA, Brown KL, Rudd KE, Buttner MJ: Analysis of the Streptomyces coelicolor sigF gene reveals the existence of a subfamily of eubacterial RNA polymerase σ factors involved in the regulation of extracytoplasmic functions. Proc Natl Acad Sci USA 1994;91:7573–7577.
  15. Maeda H, Jishage M, Nomura T, Fujita N, Ishihama A: Two extracytoplasmic function sigma subunits, σE and σFecI of Escherichia coli: promoter selectivity and intracellular levels. J Bacteriol 2000;182:1181–1184.
  16. Marx CJ, Lidstrom ME: Broad-host-range cre-lox system for antibiotic marker recycling in Gram-negative bacteria. Biotechniques 2002;33:1062–1067.
  17. Mergeay M: Bacteria adapted to industrial biotopes: metal-resistant Ralstonia; in Storz G, Hengge-Aronis G (eds): Bacterial Stress Responses. Washington, ASM Press, 2000.
  18. Mergeay M, Monchy S, Vallaeys T, Auquier V, Benotmane A, Bertin P, Taghavi S, Dunn J, van der Lelie D, Wattiez R: Ralstonia metallidurans, a bacterium specifically adapted to toxic metals: towards a catalogue of metal-responsive genes. FEMS Microbiol Rev 2003;27:385–410.
  19. Mergeay M, Nies D, Schlegel HG, Gerits J, Charles P, van Gijsegem F: Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals. J Bacteriol 1985;162:328–334.
  20. Missiakas D, Raina S: The extracytoplasmic function sigma factors: role and regulation. Mol Microbiol 1998;28:1059–1066.
  21. Mittenhuber G: An inventory of genes encoding RNA polymerase sigma factors in 31 completely sequenced eubacterial genomes. J Mol Micro Biotechnol 2002;4:77–91.
  22. Monchy S, Benotmane MA, Wattiez R, van Aelst S, Auquier V, Borremans B, Mergeay M, Taghavi S, van der Lelie D, Vallaeys T: Transcriptomics and proteomic analysis of the pMOL30-encoded copper resistance in Cupriavidus metallidurans strain CH34. Microbiology 2006;152:1765–1776.
  23. Murakami KS, Masuda S, Campbell EA, Muzzin O, Darst SA: Structural basis of transcription initiation: an RNA polymerase holoenzyme-DNA complex. Science 2002;296:1285–1290.
  24. Nies DH: CzcR and CzcD, gene products affecting regulation of resistance to cobalt, zinc and cadmium (czc system) in Alcaligenes eutrophus. J Bacteriol 1992;174:8102–8110.
  25. Nies DH: Heavy metal resistant bacteria as extremophiles: molecular physiology and biotechnological use of Ralstonia spec. CH34. Extremophiles 2000;4:77–82.
  26. Nies DH: Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol Rev 2003;27:313–339.
  27. Nies DH: Incidence and function of sigma factors in Ralstonia metallidurans and other bacteria. Arch Microbiol 2004;181:255–268.
  28. Nies DH, Brown N: Two-component systems in the regulation of heavy metal resistance; in Silver S, Walden W (eds): Metal Ions in Gene Regulation. London, Chapman Hall, 1998.
  29. Nies D, Mergeay M, Friedrich B, Schlegel HG: Cloning of plasmid genes encoding resistance to cadmium, zinc, and cobalt in Alcaligenes eutrophus CH34. J Bacteriol 1987;169:4865–4868.
  30. Nies DH, Rehbein G, Hoffmann T, Baumann C, Groβe C: Paralog of genes encoding metal resistance proteins in Cupriavidus metallidurans strain CH34. J Mol Micobiol Biotechnol 2006;147:82–93.

    External Resources

  31. Pardee AB, Jacob F, Monod J: The genetic control and cytoplasmic expression of inducibility in the synthesis of β-galactosidase of Escherichia coli. J Mol Biol 1959;1:165–168.
  32. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning, a Laboratory Manual, ed 2. Cold Spring Harbor, Cold Spring Harbor Laboratory, 1989.
  33. Shepherd NS, Dennis P, Bremer E: Cytoplasmic RNA polymerase in Escherichia coli. J Bacteriol 2001;138:2527–2534.

    External Resources

  34. Simon R, Priefer U, Pühler A: A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Biotechnology 1983;1:784–791.
  35. Stiefel A, Mahren S, Ochs M, Schindler PT, Enz S, Braun V: Control of the ferric citrate transport system of Escherichia coli: mutations in region 2.1 of the FecI extracytoplasmic-function sigma factor suppress mutations in the FecR transmembrane regulatory protein. J Bacteriol 2001;183:162–170.
  36. Tibazarwa C, Wuertz S, Mergeay M, Wyns L, van der Lelie D: Regulation of the cnr cobalt and nickel resistance determinant of Ralstoniaeutropha(Alcaligenes eutrophus) CH34. J Bacteriol 2000;182:1399–1409.
  37. Ullmann A: One-step purification of hybrid proteins which have β-galactosidase activity. Gene 1984;29:27–31.
  38. van der Lelie D, Schwuchow T, Schwidetzky U, Wuertz S, Baeyens W, Mergeay M, Nies DH: Two component regulatory system involved in transcriptional control of heavy metal homoeostasis in Alcaligenes eutrophus. Mol Microbiol 1997;23:493–503.
  39. Vandamme P, Coenye T: Taxonomy of the genus Cupriavidus: a tale of lost and found. Int J Syst Evol Microbiol 2004;54:2285–2289.
  40. Vaneechoutte M, Kämpfer P, De Baere T, Falsen E, Verschraegen G: Wautersia gen. nov., a novel genus accommodating the phylogenetic lineage including Ralstonia eutropha and related species, and proposal of Ralstonia[Pseudomonas]syzygii (Roberts et al. 1990) comb. nov. Int J Syst Evol Microbiol 2004;54:317–327.
  41. Visca P, Leoni L, Wilson MJ, Lamont IL: Iron transport and regulation, cell signalling and genomics: lessons from Escherichia coli and Pseudomonas. Mol Microbiol 2002;45:1177–1190.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50