Abstract
In order to determine the role of multidrug resistance proteins in mercury and zinc resistance, human MDR1, Lactococcus lactis lmrA, and Oenococcus oeni omrA genes were expressed in an Escherichia coli tolC mutant which is hypersensitive to metals. The three transporters conferred an increased mercury and zinc resistance to E. coli as compared to the control bacteria. This improved resistance correlated with a decreased zinc and mercury bioaccumulation. Indeed, quantification of intracellular metal concentrations by atomic absorption spectrometry (AAS) showed a 2.1-, 3-, and 5.1-fold decrease in zinc in cells expressing hMDR1, omrA, and lmrA, respectively, and a 2.7-, 7.5-, and 7.7-fold decrease in mercury in cells expressing omrA, lmrA, and hMDR1, respectively, as compared to the control bacteria. This means that hMDR1, LmrA, and OmrA proteins which are specialised in xenobiotic scavenging, their main known function, are nevertheless able to confer some resistance against metals. Our results show that the tolC mutated strain is well adapted to the study of MDR transporter activity and could be used to screen substrates and competitive hMDR1 inhibitors.
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Abbreviations
- AAS:
-
atomic absorption spectroscopy
- ABC transporter:
-
ATP binding cassette transporter
- Cd:
-
cadmium
- Hg:
-
mercury
- LB medium:
-
Luria Bertani broth medium
- GSH:
-
glutathione
- MDR:
-
multidrug resistance
- MIC:
-
minimal inhibitory concentration
- OD660 :
-
optical density at 660 nm
- Zn:
-
zinc
References
Achard M, Baudrimont M, Boudou A, Bourdineaud JP (2004) Induction of a multixenobiotic resistance protein (MXR) in the Asiatic clam Corbicula fluminea after heavy metals exposure. Aquat Toxicol 67: 347–357
Achard-Joris M, van den Berg van Saparoea HB, Driessen AJM, Bourdineaud JP. 2005 Heterologously expressed bacterial and human multidrug resistance proteins confer cadmium resistance to Escherichia coli. Biochemistry 44, 5916–5922
Ambudkar SV, Dey S, Hrycyna CA, Ramachandra M, Pastan I, Gottesman MM (1999) Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Ann Rev Pharmacol Toxicol 39: 361–398
Andrès S, Baudrimont M, Lapaquellerie Y, Ribeyre F, Maillet N, Latouche C, Boudou A (1999) Field transplantation of the fresh water bivalve Corbicula fluminea along a polymetallic contamination gradient (river Lot, France): I. Geochemical characteristics of the sampling site and cadmium and zinc bioaccumulation kinetics. Environ Toxicol Chem 18: 2462–2471
Austin EA, Graves JF, Hite LA, Parker CT, Schnaitman CA (1990) Genetic analysis of lipopolysaccharide core biosynthesis by Escherichia coli K-12: insertion mutagenesis of the rfa locus. J Bacteriol 172: 5312–5325
Baum C, Hegewisch-Becker S, Eckert HG, Stocking C, Ostertag W (1995) Novel retroviral vectors for efficient expression of the multidrug resistance (mdr-1) gene in early hematopoietic cells. J Virol 69: 7541–7547
Bourdineaud JP, Nehme B, Tesse S, Lonvaud-Funel A (2004) A bacterial gene homologous to ABC transporters protect Oenococcus oeni from ethanol and other stress factors in wine. Int J Food Microbiol 92: 1–14
Broeks A, Gerrard B, Allikmets R, Dean M, Plasterk RH (1996) Homologues of the human multidrug resistance genes MRP and MDR contribute to heavy metal resistance in the soil nematode Caenorhabditis elegans. EMBO J 15: 6132–6143
Chin KV, Tanaka S, Darlington G, Pastan I, Gottesman MM (1990) Heat shock and arsenite increase expression of the multidrug resistance (MDR1) gene in human renal carcinoma cells. J Biol Chem 265: 221–226
Dey S, Ouellette M, Lightbody J, Papadopoulou B, Rosen BP (1996) An ATP-dependent As(III)-glutathione transport system in membrane vesicles of Leishmania tarentolae. Proc Natl Acad Sci USA 93: 2192–2197
Eufemia NA, Epel D (2000) Induction of the multixenobiotic defense mechanism (MXR), P-glycoprotein, in the mussel Mytilus californianus as a general cellular response to environmental stresses. Aquat Toxicol 49: 89–100
Goldberg M, Pribyl T, Juhnke S, Nies DH (1999) Energetics and topology of CzcA, a cation/proton antiporter of the RND protein family. J Biol Chem 274: 26065–26070
Grass G, Fan B, Rosen BP, Franke S, Nies DH, Rensing C (2001) ZitB (YbgR), a member of the cation diffusion facilitator family, is an additional zinc transporter in Escherichia coli. J Bacteriol 183: 4664–4667
Hou Z-J, Narindrasorasak S, Bhushan B, Sarkar B, Mitra B (2001) Functional analysis of chimeric proteins of the Wilson Cu(I)-ATPase (ATP7B) and ZntA, a Pb(II)/Zn(II)/Cd(II)-ATPase from Escherichia coli. J Biol Chem 276: 40858–40863
Li ZS, Lu YP, Zhen RG, Szczypka M, Thiele DJ, Rea PA (1997) A new pathway for vacuolar cadmium sequestration in Saccharomyces cerevisiae: YCF1-catalyzed transport of bis(glutathionato)cadmium. Proc Natl Acad Sci USA 94: 42–47
Nies DH (2003) Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol Rev 27: 313–339
Palmiter RD, Findley SD (1995) Cloning and functional characterization of a mammalian zinc transporter that confers resistance to zinc. EMBO J 14: 639–649
Pleban K, Kopp S, Csaszar E, Peer M, Hrebicek T, Rizzi A, Ecker GF, Chiba P (2005) P-glycoprotein substrate binding domains are located at the transmembrane domain/transmembrane domain interfaces: a combined photoaffinity labeling-protein homology modeling approach. Mol Pharmacol 67: 365–374
Sharma R, Rensing C, Rosen BP, Mitra B (2000) The ATP hydrolytic activity of purified ZntA, a Pb(II)/Cd(II)/Zn(II)-translocating ATPase from Escherichia coli. J Biol Chem 275: 3873–3878
Thévenod F, Friedmann JM, Katsen AD, Hauser IA (2000) Up-regulation of multidrug resistance P-glycoprotein via nuclear factor-kappaB activation protects kidney proximal tubule cells from cadmium- and reactive oxygen species-induced apoptosis. J Biol Chem 275: 1887–1896
Tisa LS, Sekelsky JJ, Adler J (2000) Effects of organic antagonists of Ca2+, Na+, and K+ on chemotaxis and motility of Escherichia coli. J Bacteriol 182: 4856–4861
Tommasini R, Evers R, Vogt E, Mornet C, Zaman GJ, Schinkel AH, Borst P, Martinoia E (1996) The human multidrug resistance-associated protein functionally complements the yeast cadmium resistance factor 1. Proc Natl Acad Sci USA 93: 6743–6748
Tran D, Bourdineaud JP, Massabuau JC, Garnier-Laplace J (2005) Modulation of uranium bioaccumulation by hypoxia in the freshwater clam Corbicula fluminea: induction of MXR and Hsp60 in gill tissues. Environ Toxicol Chem 24: 2278–2284
van Veen HW, Venema K, Bolhuis H, Oussenko I, Kok J, Poolman B, Driessen AJM, Konings WM (1996) Multidrug resistance mediated by a bacterial homolog of the human multidrug transporter MDR1. Proc Natl Acad Sci USA 93: 10668–10672
van Veen HW, Callaghan R, Soceneantu L, Sardini A, Konings WN, Higgins CF (1998) A bacterial antibiotic-resistance gene that complements the human multidrug-resistance P-glycoprotein gene. Nature 391: 291–295
Vernhet L, Courtois A, Allain N, Payen L, Anger JP, Guillouzo A, Fardel O (1999) Overexpression of the multidrug resistance-associated protein (MRP1) in human heavy metal-selected tumor cells. FEBS Lett 443: 321–325
Waalkes MP, Diwan BA, Weghorst CM, Bare RM, Ward JM, Rice JM (1991) Anticarcinogenic effects of cadmium in B6C3F1 mouse liver and lung. Toxicol Appl Pharmacol 110: 327–335
Acknowledgements
We would like to thank Wil Konings for the gift of pGK13 and pGKLmrA plasmids and E. coli tolC strain. We also thank Régine Maury-Brachet for her help in metal determination.
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Achard-Joris, M., Bourdineaud, JP. Heterologous expression of bacterial and human multidrug resistance proteins protect Escherichia coli against mercury and zinc contamination. Biometals 19, 695–704 (2006). https://doi.org/10.1007/s10534-006-9006-2
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DOI: https://doi.org/10.1007/s10534-006-9006-2