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. 1996 Dec;178(24):7241–7247. doi: 10.1128/jb.178.24.7241-7247.1996

Identification of a third secondary carrier (DcuC) for anaerobic C4-dicarboxylate transport in Escherichia coli: roles of the three Dcu carriers in uptake and exchange.

E Zientz 1, S Six 1, G Unden 1
PMCID: PMC178639  PMID: 8955408

Abstract

In Escherichia coli, two carriers (DcuA and DcuB) for the transport of C4 dicarboxylates in anaerobic growth were known. Here a novel gene dcuC was identified encoding a secondary carrier (DcuC) for C4 dicarboxylates which is functional in anaerobic growth. The dcuC gene is located at min 14.1 of the E. coli map in the counterclockwise orientation. The dcuC gene combines two open reading frames found in other strains of E. coli K-12. The gene product (DcuC) is responsible for the transport of C4 dicarboxylates in DcuA-DcuB-deficient cells. The triple mutant (dcuA dcuB dcuC) is completely devoid of C4-dicarboxylate transport (exchange and uptake) during anaerobic growth, and the bacteria are no longer capable of growth by fumarate respiration. DcuC, however, is not required for C4-dicarboxylate uptake in aerobic growth. The dcuC gene encodes a putative protein of 461 amino acid residues with properties typical for secondary procaryotic carriers. DcuC shows sequence similarity to the two major anaerobic C4-dicarboxylate carriers DcuA and DcuB. Mutants producing only DcuA, DcuB, or DcuC were prepared. In the mutants, DcuA, DcuB, and DcuC were each able to operate in the exchange and uptake mode.

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Selected References

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  1. Alting-Mees M. A., Short J. M. pBluescript II: gene mapping vectors. Nucleic Acids Res. 1989 Nov 25;17(22):9494–9494. doi: 10.1093/nar/17.22.9494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen W. P., Kuo T. T. A simple and rapid method for the preparation of gram-negative bacterial genomic DNA. Nucleic Acids Res. 1993 May 11;21(9):2260–2260. doi: 10.1093/nar/21.9.2260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Engel P., Krämer R., Unden G. Anaerobic fumarate transport in Escherichia coli by an fnr-dependent dicarboxylate uptake system which is different from the aerobic dicarboxylate uptake system. J Bacteriol. 1992 Sep;174(17):5533–5539. doi: 10.1128/jb.174.17.5533-5539.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Engel P., Krämer R., Unden G. Transport of C4-dicarboxylates by anaerobically grown Escherichia coli. Energetics and mechanism of exchange, uptake and efflux. Eur J Biochem. 1994 Jun 1;222(2):605–614. doi: 10.1111/j.1432-1033.1994.tb18903.x. [DOI] [PubMed] [Google Scholar]
  5. Engelke T., Jording D., Kapp D., Pühler A. Identification and sequence analysis of the Rhizobium meliloti dctA gene encoding the C4-dicarboxylate carrier. J Bacteriol. 1989 Oct;171(10):5551–5560. doi: 10.1128/jb.171.10.5551-5560.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fleischmann R. D., Adams M. D., White O., Clayton R. A., Kirkness E. F., Kerlavage A. R., Bult C. J., Tomb J. F., Dougherty B. A., Merrick J. M. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science. 1995 Jul 28;269(5223):496–512. doi: 10.1126/science.7542800. [DOI] [PubMed] [Google Scholar]
  7. Fujita N., Mori H., Yura T., Ishihama A. Systematic sequencing of the Escherichia coli genome: analysis of the 2.4-4.1 min (110,917-193,643 bp) region. Nucleic Acids Res. 1994 May 11;22(9):1637–1639. doi: 10.1093/nar/22.9.1637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gutowski S. J., Rosenberg H. Succinate uptake and related proton movements in Escherichia coli K12. Biochem J. 1975 Dec;152(3):647–654. doi: 10.1042/bj1520647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
  10. Kay W. W., Kornberg H. L. The uptake of C4-dicarboxylic acids by Escherichia coli. Eur J Biochem. 1971 Jan;18(2):274–281. doi: 10.1111/j.1432-1033.1971.tb01240.x. [DOI] [PubMed] [Google Scholar]
  11. Kleckner N., Bender J., Gottesman S. Uses of transposons with emphasis on Tn10. Methods Enzymol. 1991;204:139–180. doi: 10.1016/0076-6879(91)04009-d. [DOI] [PubMed] [Google Scholar]
  12. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  13. Lambden P. R., Guest J. R. Mutants of Escherichia coli K12 unable to use fumarate as an anaerobic electron acceptor. J Gen Microbiol. 1976 Dec;97(2):145–160. doi: 10.1099/00221287-97-2-145. [DOI] [PubMed] [Google Scholar]
  14. Lo T. C. The molecular mechanism of dicarboxylic acid transport in Escherichia coli K 12. J Supramol Struct. 1977;7(3-4):463–480. doi: 10.1002/jss.400070316. [DOI] [PubMed] [Google Scholar]
  15. Needleman S. B., Wunsch C. D. A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol. 1970 Mar;48(3):443–453. doi: 10.1016/0022-2836(70)90057-4. [DOI] [PubMed] [Google Scholar]
  16. Six S., Andrews S. C., Unden G., Guest J. R. Escherichia coli possesses two homologous anaerobic C4-dicarboxylate membrane transporters (DcuA and DcuB) distinct from the aerobic dicarboxylate transport system (Dct). J Bacteriol. 1994 Nov;176(21):6470–6478. doi: 10.1128/jb.176.21.6470-6478.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sofia H. J., Burland V., Daniels D. L., Plunkett G., 3rd, Blattner F. R. Analysis of the Escherichia coli genome. V. DNA sequence of the region from 76.0 to 81.5 minutes. Nucleic Acids Res. 1994 Jul 11;22(13):2576–2586. doi: 10.1093/nar/22.13.2576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tatusov R. L., Mushegian A. R., Bork P., Brown N. P., Hayes W. S., Borodovsky M., Rudd K. E., Koonin E. V. Metabolism and evolution of Haemophilus influenzae deduced from a whole-genome comparison with Escherichia coli. Curr Biol. 1996 Mar 1;6(3):279–291. doi: 10.1016/s0960-9822(02)00478-5. [DOI] [PubMed] [Google Scholar]
  19. Unden G., Becker S., Bongaerts J., Schirawski J., Six S. Oxygen regulated gene expression in facultatively anaerobic bacteria. Antonie Van Leeuwenhoek. 1994;66(1-3):3–22. doi: 10.1007/BF00871629. [DOI] [PubMed] [Google Scholar]
  20. Van der Rest M. E., Abee T., Molenaar D., Konings W. N. Mechanism and energetics of a citrate-transport system of Klebsiella pneumoniae. Eur J Biochem. 1991 Jan 1;195(1):71–77. doi: 10.1111/j.1432-1033.1991.tb15677.x. [DOI] [PubMed] [Google Scholar]
  21. Wallace B. J., Young I. G. Role of quinones in electron transport to oxygen and nitrate in Escherichia coli. Studies with a ubiA- menA- double quinone mutant. Biochim Biophys Acta. 1977 Jul 7;461(1):84–100. doi: 10.1016/0005-2728(77)90071-8. [DOI] [PubMed] [Google Scholar]
  22. Wright J. K., Seckler R., Overath P. Molecular aspects of sugar:ion cotransport. Annu Rev Biochem. 1986;55:225–248. doi: 10.1146/annurev.bi.55.070186.001301. [DOI] [PubMed] [Google Scholar]
  23. Yura T., Mori H., Nagai H., Nagata T., Ishihama A., Fujita N., Isono K., Mizobuchi K., Nakata A. Systematic sequencing of the Escherichia coli genome: analysis of the 0-2.4 min region. Nucleic Acids Res. 1992 Jul 11;20(13):3305–3308. doi: 10.1093/nar/20.13.3305. [DOI] [PMC free article] [PubMed] [Google Scholar]

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