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Sequence analysis and interposon mutagenesis of a sensor-kinase (DctS) and response-regulator (DctR) controlling synthesis of the high-affinity C4-dicarboxylate transport system in Rhodobacter capsulatus


A two-component sensor-regulator system has been identified in the purple photosynthetic bacterium Rhodobacter capsulatus, which controls the expression of high-affinity C4-dicarboxylate transport activity in these cells. Nucleotide sequencing has revealed the existence of two genes, dctS and dctR, which together form an operon linked to, but divergently transcribed from, the previously identified dctP gene, which encodes the periplasmic binding protein of the transport system. The DctS protein is predicted to be a membrane-bound sensor-kinase with two potential membrane-spanning sequences in the N-terminal region. DctR was found to have sequence similarity throughout its entire length with proteins in the FixJ subfamily of response-regulators, especially to FixJ itself (42% identical residues). Insertional inactivation of the dctS and dctR genes resulted in the inability of the resulting mutants to grow on or transport malate, succinate or fumarate under aerobic conditions in the dark, and such mutants did not express the DctP protein. The mutants were complemented in trans by plasmids containing intact copies of the dctS and dctR genes.

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  1. Akrigg D, Bleasby AJ, Dix NIM, Findlay JBC, North ACT, Parry-Smith D, Wooton JC, Blundell TL, Gardner S, Hayes F, Islam S, Sternberg MJE, Thornton JM, Tickle IJ, Murray-Rust P (1988) A protein sequence/structure database. Nature 335:745–746

  2. Ambulos NP Jr, Smith T, Mulbury W, Lovett PS (1990) CUG as a mutant start codon for cat-86 and xylE in Bacillus subtilis. Gene 94:125–128

  3. Armstrong G, Alberti M, Leach F, Hearst JE (1989) Nucleotide sequence, organisation and nature of the protein products of the carotenoid biosynthesis gene cluster of Rhodobacter capsulatus. Mol Gen Genet 216:254–268

  4. Batut J, Santero E, Kustu S (1991) In vitro activity of the nitrogen fixation regulatory protein FixJ from Rhizobium meliloti. J Bacteriol 173:5914–5917

  5. David M, Daveran ML, Batut J, Dedieu A, Domergue O, Ghai J, Hertig C, Boistard P, Khan D (1988) Cascade regulation of nif gene expression in Rhizobium meliloti. Cell 54:671–683

  6. Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad hostrange DNA cloning system for Gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351

  7. Dodd IB, Eagan JB (1990) Improved detection of helix-turn-helix DNA binding motifs in protein sequences. Nucleic Acids Res 18:5019–5026

  8. Engelke T, Jording D, Kapp D, Pühler A (1989) Identification and sequence analysis of the Rhizobium meliloti dctA gene encoding the C4-dicarboxylate carrier. J Bacteriol 171:5551–5560

  9. Foster-Hartnett D, Kranz RG (1992) Analysis of the promoters and upstream sequences of nifA1 and nifA2 in Rhodobacter capsulatus; activation requires ntrC but not rpoN. Mol Microbiol 6:1049–1060

  10. Gibson J (1975) Uptake of C4-dicarboxylates and pyruvate by Rhodopseudomonas sphaeroides. J Bacteriol 123:471–480

  11. Gilles-Gonzalez MA, Ditta G, Helinski DR (1991) A haemoprotein with kinase activity encoded by the oxygen sensor of Rhizobium meliloti. Nature 350:170–172

  12. Göttfert M, Grob P, Hennecke H (1990) Proposed regulatory pathway encoded by the nodV and nodaW genes, determinants of host specificity in Bradyrhizobium japonicum. Proc Natl Acad Sci 87:2680–2684

  13. Hamblin MI, Shaw JG, Curson JP, Kelly DJ (1990) Mutagenesis, cloning and complementation analysis of C4-dicarboxylate transport genes from Rhodobacter capsulatus. Mol Microbiol 4:1567–1574

  14. Hillmer P, Gest H (1977) H2 metabolism in the photosynthetic bacterium Rhodopseudomonas capsulata: H2 production by growing cultures. J Bacteriol 129:724–731

  15. Jones R, Haselkorn R (1989) The DNA sequence of the Rhodobacter capsulatus ntrA, ntrB and ntrC gene analogues required for nitrogen fixation. Mol Gen Genet 215:507–516

  16. Kelly DJ, Hamblin MJ, Shaw JG (1990) Physiology and genetics of C4-dicarboxylate transport in Rhodobacter capsulatus. In: Drews G, Dawes EA (eds) Molecular biology of membrane-bound complexes in phototrophic bacteria (FEMS Symposium 53). Plenum Press, pp 453–462

  17. Khan D, Ditta G (1991) Modular structure of FixJ: homology of the transcriptional activator domain with the — 35 domain of bacterial sigma factors. Mol Microbiol 5:987–997

  18. Kranz RG, Haselkorn R (1985) Characterisation of nif regulatory genes in Rhodopseudomonas capsulata using lac gene fusions. Gene 40:203–215

  19. Kranz RG, Foster-Hartnett D (1990) Transcriptional regulatory cascade of nitrogen-fixation genes in anoxygenic photosynthetic bacteria: oxygen and nitrogen-responsive factors. Mol Microbiol 4:1793–1800

  20. Kyte J, Doolittle RF (1982) A simple method for displaying the hydrophathic character of a protein. J Mol Biol 157:105–132

  21. MacGregor BJ, Donohue TJ (1991) Evidence for two promoters for the cytochrome c 2 gene (cycA) of Rhodobacter sphaeroides. J Bacteriol 173:3949–3957

  22. Parke D (1990) Construction of mobilizable vectors derived from plasmids RP4, pUC18 and pUC19. Gene 93:135–137

  23. Prentki P, Krisch HM (1984) In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29:303–313

  24. Ronson CW, Astwood PM, Nixon BT, Ausubel FM (1987a) Deduced products of C4-dicarboxylate transport regulatory genes of Rhizobium leguminosarum are homologous to nitrogen regulatory gene products. Nucleic Acids Res 15:7921–7934

  25. Ronson CW, Nixon BT, Ausubel FM (1987b) Conserved domains in bacterial regulatory proteins that respond to environmental stimuli. Cell 49:571–581

  26. Ronson CW, Nixon BT, Albright LM, Ausubel FM (1987c) Rhizobium meliloti ntrA (rpoN) gene is required for diverse metabolic functions. J Bacteriol 169:2424–2431

  27. Rouch DA, Cram DS, DiBerardino D, Littlejohn TG, Skurray RA (1990) Efflux-mediated antiseptic resistance gene qacA from Staphylococcus aureus: common ancestry with tetracycline- and sugar-transport proteins. Mol Microbiol 4:2051–2062

  28. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

  29. Shaw JG, Kelly DJ (1991) Binding protein-dependent transport of C4-dicarboxylates in Rhodobacter capsulatus. Arch Microbiol 155:466–472

  30. Shaw JG, Hamblin MJ, Kelly DJ (1991) Purification, characterisation and nucleotide sequence of the periplasmic C4-dicarboxylate binding protein (DctP) from Rhodobacter capsulatus. Mol Microbiol 5:3055–3062

  31. Staden R (1980) A new computer method for the storage and manipulation of DNA gel reading data. Nucleic Acids Res 8:3673–3694

  32. Staden R (1982) An interactive graphics program for comparing and aligning nucleic acid and amino-acid sequences. Nucleic Acids Res 10:2951–2961

  33. Staden R, McLachlan AD (1982) Codon preference and its use in identifying protein coding regions in long DNA stretches. Nucleic Acids Res 10:141–156

  34. Stahl CL, Scjka GA (1973) Growth of Rhodopseudomonas capsulata on l- and d-malic acid. Biochim Biophys Acta 299:241–245

  35. Stock JB, Ninfa AJ, Stock AM (1989) Protein phosphorylation and the regulation of adaptive responses in bacteria. Microbiol Rev 53:450–490

  36. Taylor DP, Cohen SN, Clark WG, Marrs BL (1983) Alignment of the genetic and restriction maps of the photosynthesis regions of the Rhodopseudomonas capsulata chromosome by a conjugation-mediated marker rescue technique. J Bacteriol 154:580–590

  37. Walmsley AR, Shaw JG, Kelly DJ (1992) The mechanism of ligand binding to the periplasmic C4-dicarboxylate binding protein (DctP) from Rhodobacter capsulatus. J Biol Chem 267:8064–8072

  38. Weaver PF, Wall JD, Gest H (1975) Characterisation of Rhodopseudomonas capsulata. Arch Microbiol 105:207–216

  39. Weston LA, Kadner RJ (1988) Role of uhp genes in expression of the Escherichia coli sugar-phosphate transport system. J Bacteriol 170:3375–3383

  40. Woese CR (1987) Bacterial evolution. Microbiol Rev 51:221–271

  41. Wootton JC, Drummond MH (1989) The Q-linker: a class of interdomain sequences found in bacterial multidomain regulatory proteins. Protein Eng 2:535–543

  42. Yarosh OK, Charles TC, Finan TM (1989) Analysis of C4-dicarboxylate transport genes in Rhizobium meliloti. Mol Microbiol 3:813–823

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Correspondence to David J. Kelly.

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Communicated by J.W. Lengeler

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Hamblin, M.J., Shaw, J.G. & Kelly, D.J. Sequence analysis and interposon mutagenesis of a sensor-kinase (DctS) and response-regulator (DctR) controlling synthesis of the high-affinity C4-dicarboxylate transport system in Rhodobacter capsulatus . Molec. Gen. Genet. 237, 215–224 (1993). https://doi.org/10.1007/BF00282803

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Key words

  • Sensor-kinase
  • Response-regulator
  • C4-dicarboxylates
  • Transport
  • Rhodobacter capsulatus