Abstract
The transporter RcnA has previously been implicated in Ni(II) and Co(II) detoxification in E. coli probably through efflux. Here we demonstrate that the divergently described rcnA and rcnR gene products constitute a link between nickel, cobalt and iron homeostasis. Deletion of the rcnA gene resulted in increased cellular nickel, cobalt and iron concentrations. Expression of rcnA was induced by Ni(II) or Co(II). Overproduction of rcnR inhibited induction of rcnA by metal cations but RcnR did not bind to the rcnA promoter in vitro. When rcnR or fur, the gene of the global repressor of iron homeostasis, was deleted, expression of rcnA was also induced by iron. The promoter region of rcnA was positive in a Fur titration (FURTA) in vivo assay indicative of Fur binding. Thus, rcnA is part of the Fur regulon of E.␣coli. The implications of a connection between the homoeostasis of closely related transition metals are discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adrait A, Jacquamet L, Le Pape L, et al. (1999) Spectroscopic and saturation magnetization properties of the manganese- and cobalt-substituted fur (ferric uptake regulation) protein from Escherichia coli. Biochemistry 38:6248
Altschul SF, Madden TL, Schaffer AA, et al. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acid Res 25:3389–3402
Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557
Baichoo N, Helmann JD (2002) Recognition of DNA by Fur: a reinterpretation of the Fur box consensus sequence. J Bacteriol 184:5826–5832
Contreras M, Thiberge JM, Mandrand-Berthelot MA, Labigne A (2003) Characterization of the roles of NikR, a nickel-responsive pleiotropic autoregulator of Helicobacter pylori. Mol Microbiol 49:947–963
Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97:6640–6645
De Pina K, Desjardin V, Mandrand-Berthelot MA, Giordano G, Wu LF (1999) Isolation and characterization of the nikR gene encoding a nickel-responsive regulator in Escherichia coli. J Bacteriol 181:670–674
Delany I, Ieva R, Soragni A, Hilleringmann M, Rappuoli R, Scarlato V (2005) In vitro analysis of protein-operator interactions of the NikR and Fur metal-responsive regulators of coregulated genes in Helicobacter pylori. J Bacteriol 187:7703–7715
Ernst FD, Kuipers EJ, Heijens A, et al. (2005) The nickel-responsive regulator NikR controls activation and repression of gene transcription in Helicobacter pylori. Infect Immun 73:7252–7258
Grass G, Fan B, Rosen BP, Lemke K, Schlegel HG, Rensing C (2001) NreB from Achromobacter xylosoxidans 31A is a nickel-induced transporter conferring nickel resistance. J Bacteriol 183:2803–2807
Grass G, Franke S, Taudte N, et al. (2005a) The metal permease ZupT from Escherichia coli is a transporter with a broad substrate spectrum. J Bacteriol 187:1604–1611
Grass G, Otto M, Fricke B, et al. (2005b) FieF (YiiP) from Escherichia coli mediates decreased cellular accumulation of iron and relieves iron stress. Arch Microbiol 183:9–18
Hantke K (1987) Selection procedure for deregulated iron transport mutants (fur) in Escherichia coli K 12: fur not only affects iron metabolism. Mol Gen Genetics 210:135–139
Herring CD, Blattner FR (2004) Global transcriptional effects of a suppressor tRNA and the inactivation of the regulator frmR. J Bacteriol 186:6714–6720
Hmiel SP, Snavely MD, Florer JB, Maguire ME, Miller CG (1989) Magnesium transport in Salmonella typhimurium: genetic characterization and cloning of three magnesium transport loci. J Bacteriol 171:4742–4751
Housecroft CE, Constable EC (2006) Chemistry, 3rd edn. Essex, England, Pearson Education Limited
Ikeda JS, Janakiraman A, Kehres DG, Maguire ME, Slauch JM (2005) Transcriptional regulation of sitABCD of Salmonella enterica serovar Typhimurium by MntR and Fur. J Bacteriol 187:912
Jacquamet L, Aberdam D, Adrait A, Hazemann JL, Latour JM, Michaud-Soret I (1998) X-ray absorption spectroscopy of a new zinc site in the Fur protein from Escherichia coli. Biochemistry-USA 37:2564–2571
Marrero, J, Auling, G, Coto, O, Nies, DH. 2006 High-level resistance to cobalt and nickel but probably no transenvelope efflux: metal resistance in the Cuban Serratia marcescens strain C-1. Microb Ecol doi: 10.1007/s00248-006-9152-7
Mergeay M, Nies D, Schlegel HG, Gerits J, Charles P, van Gijsegem F (1985) Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals. J Bacteriol 162:328–334
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor, New York, Cold Spring Harbor Press
Moncrief MB, Maguire ME (1999) Magnesium transport in prokaryotes. J Biol Inorg Chem 4:523–527
Munkelt D, Grass G, Nies DH (2004) The chromosomally encoded cation diffusion facilitator proteins DmeF and FieF from Wautersia metallidurans CH34 are transporters of broad metal specificity. J Bacteriol 186:8036–8043
Nies DH (2003) Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol Rev 27:313–339
Nies, DH. 2004 Essential and toxic effects of elements on microorganisms. In: Anke K, Ihnat M, Stoeppler M, eds. Metals and their compounds in the environment. Weinheim Wiley-VCH: p Part II.1
Outten CE, O’Halloran TV (2001) Femtomolar sensitivity of metalloregulatory proteins controlling zinc homeostasis. Science 292:2488–2492
Park JE, Young KE, Schlegel HG, Rhie HG, Lee HS (2003) Conjugative plasmid mediated inducible nickel resistance in Hafnia alvei 5-5. Int Microbiol 6:57–64
Park JE, Schlegel HG, Rhie HG, Lee HS (2004) Nucleotide sequence and expression of the ncr nickel and cobalt resistance in Hafnia alvei 5-5. Int Microbiol 7:27–34
Pohl E, Haller JC, Mijovilovich A, Meyer-Klaucke W, Garman E, Vasil ML (2003) Architecture of a protein central to iron homeostasis: crystal structure and spectroscopic analysis of the ferric uptake regulator. Mol Microbiol 47:903–915
Rodrigue A, Effantin G, Mandrand-Berthelot MA (2005) Identification of rcnA (yohM), a nickel and cobalt resistance gene in Escherichia coli. J Bacteriol 187:2912–2916
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory
Stojiljkovic I, Bäumler AJ, Hantke K (1994) Fur regulon in gram-negative bacteria. Identification and characterization of new iron-regulated Escherichia coli genes by a fur titration assay. J Mol Biol 236:531–545
Stoppel RD, Meyer M, Schlegel HG (1995) The nickel resistance determinant cloned from the enterobacterium Klebsiella oxytoca: conjugational transfer, expression, regulation and DNA homologies to various nickel-resistant bacteria. Biometals 8:70–79
Tabor S, Richardson CC (1985) A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci USA 82:1074–1078
Van Vliet AHM, Kuipers EJ, Waidner B, et al. (2001) Nickel-responsive induction of urease expression in Helicobacter pylori is mediated at the transcriptional level. Infect Immun 69:4891–4897
Weast RC (1984) CRC handbook of chemistry and physics. CRC Press, Inc., Boca Raton, Florida, USA
Zheleznova EE, Crosa JH, Brennan RG (2000) Characterization of the DNA- and metal-binding properties of Vibrio anguillarum Fur reveals conservation of a structural Zn2+ ion. J Bacteriol 182:2624–2630
Acknowledgments
This work was supported by the Land Sachsen-Anhalt (HWP 56 IF) and Deutsche Forschungsgemeinschaft (Ni262/3). We thank Grit Schleuder for skilful technical assistance. Thanks are due Klaus Hantke for the generous gift of E. coli strain H1717, Nadine Taudte for construction of strains ECA348 and ECA349 and the NARA Institute of Science and Technology (Kansai Science City, Japan) for E. coli strain BW25113 Delta nikR::kan.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Koch, D., Nies, D.H. & Grass, G. The RcnRA (YohLM) system of Escherichia coli: A connection between nickel, cobalt and iron homeostasis. Biometals 20, 759–771 (2007). https://doi.org/10.1007/s10534-006-9039-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10534-006-9039-6