Molecular Analysis of the Copper-Transporting Efflux System CusCFBA of Escherichia coli
10.1128/jb.185.13.3804-3812.2003
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

ABSTRACT The cus determinant of Escherichia coli encodes the CusCFBA proteins that mediate resistance to copper and silver by cation efflux. CusA and CusB were essential for copper resistance, and CusC and CusF were required for full resistance. Replacements of methionine residues 573, 623, and 672 with isoleucine in CusA resulted in loss of copper resistance, demonstrating their functional importance. Substitutions for several other methionine residues of this protein did not have any effect. The small 10-kDa protein CusF (previously YlcC) was shown to be a periplasmic protein. CusF bound one copper per polypeptide. The pink CusF copper protein complex exhibited an absorption maximum at around 510 nm. Methionine residues of CusF were involved in copper binding as shown by site-directed mutagenesis. CusF interacted with CusB and CusC polypeptides in a yeast two-hybrid assay. In contrast to other well-studied CBA-type heavy metal efflux systems, Cus was shown to be a tetrapartite resistance system that involves the novel periplasmic copper-binding protein CusF. These data provide additional evidence for the hypothesis that Cu(I) is directly transported from the periplasm across the outer membrane by the Cus complex.

Bibliography

Franke, S., Grass, G., Rensing, C., & Nies, D. H. (2003). Molecular Analysis of the Copper-Transporting Efflux System CusCFBA of Escherichia coli. Journal of Bacteriology, 185(13), 3804–3812.

Authors 4
  1. Sylvia Franke (first)
  2. Gregor Grass (additional)
  3. Christopher Rensing (additional)
  4. Dietrich H. Nies (additional)
References 67 Referenced 439
  1. Andersen, C., C. Hughes, and V. Koronakis. 2001. Protein export and drug efflux through bacterial channel-tunnels. Curr. Opin. Cell Biol.13:412-416. (10.1016/S0955-0674(00)00229-5) / Curr. Opin. Cell Biol. (2001)
  2. Andersen, C., E. Koronakis, C. Hughes, and V. Koronakis. 2002. An aspartate ring at the TolC tunnel entrance determines ion selectivity and presents a target for blocking by large cations. Mol. Microbiol.44:1131-1139. (10.1046/j.1365-2958.2002.02898.x) / Mol. Microbiol. (2002)
  3. Anraku, Y., and R. B. Gennis. 1987. The aerobic respiratory chain of Escherichia coli. Trends Biochem. Sci.12:262-266. (10.1016/0968-0004(87)90131-9) / Trends Biochem. Sci. (1987)
  4. Anton, A., C. Große, J. Reißman, T. Pribyl, and D. H. Nies. 1999. CzcD is a heavy metal ion transporter involved in regulation of heavy metal resistance in Ralstonia sp. strain CH34. J. Bacteriol.181:6876-6881. / strain CH34. J. Bacteriol. (1999)
  5. 10.1128/JB.180.4.938-944.1998
  6. Arnesano, F., L. Banci, I. Bertini, S. Mangani, and A. R. Thompsett. 2003. A redox switch in CopC: an intriguing copper trafficking protein that binds copper(I) and copper(II) to different sites. Proc. Natl. Acad. Sci. USA100:3814-3819. (10.1073/pnas.0636904100) / Proc. Natl. Acad. Sci. USA (2003)
  7. Berry, S. M., X. Wang, and Y. Lu. 2000. Ligand replacement study at the His120 site of purple CuA azurin. J. Inorg. Biochem.78:89-95. (10.1016/S0162-0134(99)00214-7) / J. Inorg. Biochem. (2000)
  8. Bloss, T., S. Clemens, and D. H. Nies. 2002. Characterisation of the ZAT1p zinc transporter from Arabidopsis thaliana in microbial model organisms and reconstituted proteoliposomes. Planta214:783-791. (10.1007/s00425-001-0677-1) / Planta (2002)
  9. 10.1016/0003-2697(76)90527-3
  10. Brown, N. L., S. R. Barrett, J. Camakaris, B. T. Lee, and D. A. Rouch. 1995. Molecular genetics and transport analysis of the copper-resistance determinant (pco) from Escherichia coli plasmid pRJ1004. Mol. Microbiol.17:1153-1166. (10.1111/j.1365-2958.1995.mmi_17061153.x) / Mol. Microbiol. (1995)
  11. Cooper, R. A., P. F. Knowles, D. E. Brown, M. A. McGuirl, and D. M. Dooley. 1992. Evidence for copper and 3,4,6-trihydroxyphenylalanine quinone cofactors in an amine oxidase from the Gram-negative bacterium Escherichia coli K12. Biochem. J.288:337-340. (10.1042/bj2880337) / Biochem. J. (1992)
  12. Dancis, A., D. Haile, D. S. Yuan, and R. D. Klausner. 1994. The Saccharomyces cerevisiae copper transport protein (Ctr1p). Biochemical characterization, regulation by copper, and physiologic role in copper uptake. J. Biol. Chem.269:25660-25667. (10.1016/S0021-9258(18)47300-0) / J. Biol. Chem. (1994)
  13. 10.1073/pnas.120163297
  14. 10.1128/jb.176.13.3825-3831.1994
  15. 10.1128/JB.184.23.6490-6499.2002
  16. Fan, B., G. Grass, C. Rensing, and B. P. Rosen. 2001. Escherichia coli CopA N-terminal Cys(X)(2)Cys motifs are not required for copper resistance or transport. Biochem. Biophys. Res. Commun.286:414-418. (10.1006/bbrc.2001.5367) / Biochem. Biophys. Res. Commun. (2001)
  17. Franke, S., G. Grass, and D. H. Nies. 2001. The product of the ybdE gene of the Escherichia coli chromosome is involved in detoxification of silver ions. Microbiology147:965-972. (10.1099/00221287-147-4-965) / Microbiology (2001)
  18. Goldberg, M., T. Pribyl, S. Juhnke, and D. H. Nies. 1999. Energetics and topology of CzcA, a cation/proton antiporter of the RND protein family. J. Biol. Chem.274:26065-26070. (10.1074/jbc.274.37.26065) / J. Biol. Chem. (1999)
  19. Gort, A. S., D. M. Ferber, and J. A. Imlay. 1999. The regulation and role of the periplasmic copper, zinc superoxide dismutase of Escherichia coli. Mol. Microbiol.32:179-191. (10.1046/j.1365-2958.1999.01343.x) / Mol. Microbiol. (1999)
  20. Grass G. 2000. Molekulargenetische und biochemische Charakterisierung der cnr Cobalt/Nickel-Resistenz-Determinante aus Ralstonia metallidurans CH34. Dissertation. Martin-Luther-Universität Halle-Wittenberg Halle/Saale Germany.
  21. Grass, G., and C. Rensing. 2001. CueO is a multi-copper oxidase that confers copper tolerance in Escherichia coli. Biochem. Biophys. Res. Commun.286:902-908. (10.1006/bbrc.2001.5474) / Biochem. Biophys. Res. Commun. (2001)
  22. 10.1128/JB.183.6.2145-2147.2001
  23. Gupta, A., K. Matsui, J. F. Lo, and S. Silver. 1999. Molecular basis for resistance to silver in Salmonella. Nat. Med.5:183-188. (10.1038/5545) / Nat. Med. (1999)
  24. Gupta, A., L. T. Phung, D. E. Taylor, and S. Silver. 2001. Diversity of silver resistance genes in IncH incompatibility group plasmids. Microbiology147:3393-3402. (10.1099/00221287-147-12-3393) / Microbiology (2001)
  25. Hantke, K. 2001. Bacterial zinc transporters and regulators. Biometals14:239-249. (10.1023/A:1012984713391) / Biometals (2001)
  26. Hay, M., J. H. Richards, and Y. Lu. 1996. Construction and characterization of an azurin analog for the purple copper site in cytochrome c oxidase. Proc. Natl. Acad. Sci. USA93:461-464. (10.1073/pnas.93.1.461) / Proc. Natl. Acad. Sci. USA (1996)
  27. Higgins, C. F. 1992. ABC-transporters: from microorganisms to man. Annu. Rev. Cell Biol.8:67-113. (10.1146/annurev.cb.08.110192.000435) / Annu. Rev. Cell Biol. (1992)
  28. Huffman, D. L., J. Huyett, F. W. Outten, P. E. Doan, L. A. Finney, B. M. Hoffman, and T. V. O'Halloran. 2002. Spectroscopy of Cu(II)-PcoC and the multicopper oxidase function of PcoA, two essential components of Escherichia coli pco copper resistance operon. Biochemistry41:10046-10055. (10.1021/bi0259960) / Biochemistry (2002)
  29. 10.1128/jb.179.20.6264-6270.1997
  30. 10.1128/JB.184.12.3159-3166.2002
  31. Knight, S. A., S. Labbe, L. F. Kwon, D. J. Kosman, and D. J. Thiele. 1996. A widespread transposable element masks expression of a yeast copper transport gene. Genes Dev.10:1917-1929. (10.1101/gad.10.15.1917) / Genes Dev. (1996)
  32. Kolczak, U., J. Salgado, G. Siegal, M. Saraste, and G. W. Canters. 1999. Paramagnetic NMR studies of blue and purple copper proteins. Biospectroscopy5(Suppl.):19-32. (10.1002/(SICI)1520-6343(1999)5:1<19::AID-BSPY4>3.0.CO;2-E) / Biospectroscopy (1999)
  33. 10.1038/35016007
  34. Lee, S. M., G. Grass, C. Rensing, S. R. Barrett, C. J. Yates, J. V. Stoyanov, and N. L. Brown. 2002. The Pco proteins are involved in periplasmic copper handling in Escherichia coli. Biochem. Biophys. Res. Commun.295:616-620. (10.1016/S0006-291X(02)00726-X) / Biochem. Biophys. Res. Commun. (2002)
  35. 10.1128/jb.175.3.767-778.1993
  36. Makdessi, K., J. R. Andreesen, and A. Pich. 2001. Tungstate uptake by a highly specific ABC transporter in Eubacterium acidaminophilum. J. Biol. Chem.276:24557-24564. (10.1074/jbc.M101293200) / J. Biol. Chem. (2001)
  37. Mao, W. M., M. S. Warren, D. S. Black, T. Satou, T. Murata, T. Nishino, N. Gotoh, and O. Lomovskaya. 2002. On the mechanism of substrate specificity by resistance nodulation division (RND)-type multidrug resistance pumps: the large periplasmic loops of MexD from Pseudomonas aeruginosa are involved in substrate recognition. Mol. Microbiol.46:889-901. (10.1046/j.1365-2958.2002.03223.x) / Mol. Microbiol. (2002)
  38. Maseda, H., M. Kitao, S. Eda, E. Yoshihara, and T. Nakae. 2002. A novel assembly process of the multicomponent xenobiotic efflux pump in Pseudomonas aeruginosa. Mol. Microbiol.46:677-686. (10.1046/j.1365-2958.2002.03197.x) / Mol. Microbiol. (2002)
  39. 10.1128/AAC.44.3.658-664.2000
  40. Mills, S. D., C. K. Lim, and D. A. Cooksey. 1994. Purification and characterization of CopR, a transcriptional activator protein that binds to a conserved domain (cop box) in copper-inducible promoters of Pseudomonas syringae. Mol. Gen. Genet.244:341-351. (10.1007/BF00286685) / Mol. Gen. Genet. (1994)
  41. 10.1128/JB.182.20.5864-5871.2000
  42. Murakami, S., R. Nakashima, R. Yamashita, and A. Yamaguchi. 2002. Crystal structure of bacterial multidrug efflux transporter AcrB. Nature419:587-593. (10.1038/nature01050) / Nature (2002)
  43. Navarro, C., L. F. Wu, and M. A. Mandrand Berthelot. 1993. The nik operon of Escherichia coli encodes a periplasmic binding-protein-dependent transport system for nickel. Mol. Microbiol.9:1181-1191. (10.1111/j.1365-2958.1993.tb01247.x) / Mol. Microbiol. (1993)
  44. Nies D. H. Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol. Rev. in press.
  45. Nies, D. H., A. Nies, L. Chu, and S. Silver. 1989. Expression and nucleotide sequence of a plasmid-determined divalent cation efflux system from Alcaligenes eutrophus. Proc. Natl. Acad. Sci. USA86:7351-7355. (10.1073/pnas.86.19.7351) / Proc. Natl. Acad. Sci. USA (1989)
  46. Nittis, T., G. N. George, and D. R. Winge. 2001. Yeast Sco1, a protein essential for cytochrome c oxidase function, is a Cu(I)-binding protein. J. Biol. Chem.276:42520-42526. (10.1074/jbc.M107077200) / J. Biol. Chem. (2001)
  47. Outten, F. W., D. L. Huffman, J. A. Hale, and T. V. O'Halloran. 2001. The independent cue and cus systems confer copper tolerance during aerobic and anaerobic growth in Escherichia coli. J. Biol. Chem.276:30670-30677. (10.1074/jbc.M104122200) / J. Biol. Chem. (2001)
  48. Patzer, S. I., and K. Hantke. 1998. The ZnuABC high-affinity zinc uptake system and its regulator Zur in Escherichia coli. Mol. Microbiol.28:1199-1210. (10.1046/j.1365-2958.1998.00883.x) / Mol. Microbiol. (1998)
  49. Pena, M. M. O., S. Puig, and D. J. Thiele. 2000. Characterization of the Saccharomyces cerevisiae high affinity copper transporter Ctr3. J. Biol. Chem.275:33244-33251. (10.1074/jbc.M005392200) / J. Biol. Chem. (2000)
  50. Petersen, C., and L. B. Moller. 2000. Control of copper homeostasis in Escherichia coli by a P-type ATPase, CopA, and a MerR-like transcriptional activator, CopR. Gene261:289-298. (10.1016/S0378-1119(00)00509-6) / Gene (2000)
  51. Qian, H., L. Sahlman, P. O. Eriksson, C. Hambraeus, U. Edlund, and I. Sethson. 1998. NMR solution structure of the oxidized form of MerP, a mercuric ion binding protein involved in bacterial mercuric ion resistance. Biochem. USA37:9316-9322. (10.1021/bi9803628) / Biochem. USA (1998)
  52. Rech, S., C. Wolin, and R. P. Gunsalus. 1996. Properties of the periplasmic ModA molybdate-binding protein of Escherichia coli. J. Biol. Chem.271:2557-2562. (10.1074/jbc.271.5.2557) / J. Biol. Chem. (1996)
  53. Rensing, C., B. Fan, R. Sharma, B. Mitra, and B. P. Rosen. 2000. CopA: an Escherichia coli Cu(I)-translocating P-type ATPase. Proc. Natl. Acad. Sci. USA97:652-656. (10.1073/pnas.97.2.652) / Proc. Natl. Acad. Sci. USA (2000)
  54. 10.1128/JB.181.19.5891-5897.1999
  55. 10.1128/jb.179.22.6871-6879.1997
  56. Saier, M. H., Jr., R. Tam, A. Reizer, and J. Reizer. 1994. Two novel families of bacterial membrane proteins concerned with nodulation, cell division and transport. Mol. Microbiol.11:841-847. (10.1111/j.1365-2958.1994.tb00362.x) / Mol. Microbiol. (1994)
  57. 10.1128/MMBR.64.2.354-411.2000
  58. Sambrook J. E. F. Fritsch and T. Maniatis. 1989. Molecular cloning: a laboratory manual 2nd ed. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  59. 10.1128/jb.176.22.7045-7054.1994
  60. 10.1128/jb.179.24.7875-7881.1997
  61. Stoyanov, J. V., J. L. Hobman, and N. L. Brown. 2001. CueR (YbbI) of Escherichia coli is a MerR family regulator controlling expression of the copper exporter CopA. Mol. Microbiol.39:502-511. (10.1046/j.1365-2958.2001.02264.x) / Mol. Microbiol. (2001)
  62. 10.1128/JB.184.23.6499-6507.2002
  63. Tsai, K. J., Y. F. Lin, M. D. Wong, H. H. C. Yang, H. L. Fu, and B. P. Rosen. 2002. Membrane topology of the pl258 CadA Cd(II)/Pb(II)/Zn(II)-translocating P-type ATPase. J. Bioenerg. Biomembr.34:147-156. (10.1023/A:1016085301323) / J. Bioenerg. Biomembr. (2002)
  64. Tseng, T.-T., K. S. Gratwick, J. Kollman, D. Park, D. H. Nies, A. Goffeau, and M. H. Saier, Jr. 1999. The RND superfamily: an ancient, ubiquitous and diverse family that includes human disease and development proteins. J. Mol. Microbiol. Biotechnol.1:107-125. / J. Mol. Microbiol. Biotechnol. (1999)
  65. Yoneyama, H., A. Ocaktan, M. Tsuda, and T. Nakae. 1997. The role of mex-gene products in antibiotic extrusion in Pseudomonas aeruginosa. Biochem. Biophys. Res. Commun.233:611-618. (10.1006/bbrc.1997.6506) / Biochem. Biophys. Res. Commun. (1997)
  66. Zgurskaya, H. I., and H. Nikaido. 2000. Multidrug resistance mechanisms: drug efflux across two membranes. Mol. Microbiol.37:219-225. (10.1046/j.1365-2958.2000.01926.x) / Mol. Microbiol. (2000)
  67. Zhou, H., and D. J. Thiele. 2001. Identification of a novel high affinity copper transport complex in the fission yeast Schizosaccharomyces pombe. J. Biol. Chem.276:20529-20535. (10.1074/jbc.M102004200) / J. Biol. Chem. (2001)
Dates
Type When
Created 22 years, 2 months ago (June 17, 2003, 1:59 p.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 2:09 p.m.)
Indexed 3 days, 23 hours ago (Sept. 3, 2025, 6:29 a.m.)
Issued 22 years, 2 months ago (July 1, 2003)
Published 22 years, 2 months ago (July 1, 2003)
Published Print 22 years, 2 months ago (July 1, 2003)
Funders 0

None

@article{Franke_2003, title={Molecular Analysis of the Copper-Transporting Efflux System CusCFBA of Escherichia coli}, volume={185}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.185.13.3804-3812.2003}, DOI={10.1128/jb.185.13.3804-3812.2003}, number={13}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Franke, Sylvia and Grass, Gregor and Rensing, Christopher and Nies, Dietrich H.}, year={2003}, month=jul, pages={3804–3812} }