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Volume 143, Issue 6

employs a single triosephosphate isomerase for heterotrophic and autotrophic metabolism Free

Abstract

The expression of the and operons of encoding enzymes of the Calvin cycle is regulated by the transcriptional regulator CbbR. In order to identify other genes involved in the regulation of these operons, a mutant was isolated with a lowered activity of a fusion between the promoter of the operon and the reporter gene This mutant was unable to grow autotrophically and had a reduced growth rate of medium supplemented with gluconate or succinate. The regulation of the operon in the mutant was indistinguishable from the wild-type strain, but induction of the operon upon transition to autotrophic growth conditions was delayed. Complementation of the mutant with a genomic library of resulted in the isolation of a 1∙1 kb I fragment which restored autotrophic growth of the mutant. One open reading frame (ORF) was present on the I fragment, which could encode a protein highly similar to triosephosphate isomerase proteins from other bacteria. Cell extracts of the mutant grown under glycolytic or gluconeogenic conditions had severely reduced triosephosphate isomerase activities. The ORF was therefore identified as encoding triosephosphate isomerase. The gene is not linked to the previously identified operons encoding Calvin cycle enzymes and therefore represents a third transcriptional unit required for autotrophic metabolism.

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Keyword(s): autotrophy , regulation , RuBisCO , triosephosphate isomerase and Xanthobacter flavus
© Society Society for General Microbiology 1997
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1997-06-01
2024-05-06
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References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool.. J Mol Biol 215:403–410
     [Google Scholar]
  2. van den Bergh E. R. E., Dijkhuizen L., Meijer W. G. 1993; CbbR, a LysR-type transcriptional activator, is required for expression of the autotrophic CO2 fixation enzymes of Xanthobacter flavus.. J Bacterial 175:6097–6104
     [Google Scholar]
  3. van den Bergh E. R. E., van der Kooij T. A. W., Dijkhuizen L., Meijer W. G. 1995; Fructosebisphosphatase isoenzymes of the chemoautotroph Xanthobacter flavus.. J Bacterial 177:5860–5864
     [Google Scholar]
  4. van den Bergh E. R. E., Baker S. C., Raggers R. J., Terpstra P., Woudstra E. C., Dijkhuizen L., Meijer W. G. 1996; Primary structure and phylogeny of the Calvin cycle enzymes transketolase and fructosebisphosphate aldolase of Xanthobacter flavus.. J Bacterial 178:888–893
     [Google Scholar]
  5. birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening of recombinant plasmid DNA.. Nucleic Acids Res 7:1513–1523
     [Google Scholar]
  6. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding.. Anal Biochem 73:248–254
     [Google Scholar]
  7. Chen J.-H., Gibson J. L., McCue L. A., Tabita F. R. 1991; Identification, expression and deduced primary structure of transketolase and other enzymes encoded within the form II CO2 fixation operon of Rhodobacter spbaeroides.. J Biol Chem 266:20447–20452
     [Google Scholar]
  8. Croes L. M., Meijer W. G., Dijkhuizen L. 1991; Regulation of methanol oxidation and carbon dioxide fixation in Xantbobacter strain 25a grown in continuous culture.. Arch Microbiol 155:159–163
     [Google Scholar]
  9. Dijkhuizen L., Harder W. 1984; Current views on the regulation of autotrophic carbon dioxide fixation via the Calvin cycle in bacteria.. Antonie Leeuwenhoek 50:473–487
     [Google Scholar]
  10. Ditta G., Stanfield S., Corbin D., Helinski D. R. 1980; Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti.. Proc Natl Acad Sci USA 77:7347–7351
     [Google Scholar]
  11. Falcone D. L., Tabita F. R. 1993; Complementation analysis and regulation of CO2 fixation gene expression in a ribulose 1,5-bisphosphate carboxylase-oxygenase deletion strain of Rbodo-spirillum rubrum.. J Bacterial 175:5066–5077
     [Google Scholar]
  12. gibson J. L., Tabita F. R. 1977; Different molecular forms of d-ribulose-l,5-bisphosphate carboxylase from Rbodopseudomonas spbaeroides.. J Biol Chem 252:943–949
     [Google Scholar]
  13. Gibson J. L., Tabita F. R. 1996; The molecular regulation of the reductive pentose phosphate pathway in Proteobacteria and Cyanobacteria. . Arch Microbiol 166:141–150
     [Google Scholar]
  14. Gibson J. L., Falcone D. L., Tabita F. R. 1991; Nucleotide sequence, transcriptional analysis, and expression of genes encoded within the form I CO2 fixation operon of Rhodobacter spbaeroides.. J Biol Chem 266:14646–14653
     [Google Scholar]
  15. Im D., Friedrich C. G. 1983; Fluoride, hydrogen and formate activate ribulosebisphosphate carboxylase formation in Alcali-genes eutrophus.. J Bacteriol 154:803–808
     [Google Scholar]
  16. Irani M. H., Maitra P. K. 1977; Properties of Escherichia coli mutants deficient in enzymes of glycolysis.. J Bacteriol 132:398–410
     [Google Scholar]
  17. Keen N. T., Tamaki S., Kobayashi D., Trollinger D. 1988; Improved broad-host-range plasmids for DNA cloning in Gramnegative bacteria.. Gene 70:191–197
     [Google Scholar]
  18. Knauf V. C., Nester E. W. 1982; Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid.. Plasmid 8:45–54
     [Google Scholar]
  19. Leadbeater L., Si Bowien B. 1984; Control of autotrophic carbon assimilation in Alcaligenes eutrophus by the inactivation and reactivation of phosphoribulokinase.. J Bacteriol 157:95–99
     [Google Scholar]
  20. Lehmicke L. G., Lidstrom M. E. 1985; Organization of genes necessary for growth of the hydrogen-methanol autotroph Xantbobacter sp. strain H4-14 on hydrogen and carbon dioxide.. J Bacteriol 162:1244–1249
     [Google Scholar]
  21. Lidstrom-O’Conner M. E., Fulton G. L., Wopat A. E. 1983; ‘Methylobacterium ethanolicum’: a syntrophic association of two methylotrophic bacteria.. J Gen Microbiol 129:3139–3148
     [Google Scholar]
  22. Meijer W. G. 1994; The Calvin cycle enzyme phosphoglycerate kinase of Xantbobacter flavus required for autotrophic CO2 fixation is not encoded by the cbb operon.. J Bacteriol 176:6120–6126
     [Google Scholar]
  23. Meijer W. G. 1996; Genetics of CO2 fixation in methylotrophs. In Proceedings of the 8th International Symposium on Microbial Growth on C1 Compounds, pp.. 118–125 Edited by M. E. Lidstrom & F. R. Tabita. Dordrecht: Kluwer Academic..
    [Google Scholar]
  24. Meijer W. G., Dijkhuizen L. 1988; Regulation of autotrophic metabolism in Pseudomonas oxalaticus OX1 wild-type and an isocitrate-lyase-deficient mutant.. J Gen Microbiol 134:3231–3237
     [Google Scholar]
  25. Meijer W. G., Croes L. M., Jenni B., Lehmicke L. G., Lidstrom M. E., Dijkhuizen L. 1990a; Characterization of Xantbobacter strains H4-14 and 25a and enzyme profiles after growth under autotrophic and heterotrophic growth conditions.. Arch Microbiol 153:360–367
     [Google Scholar]
  26. Meijer W. G., Enequist H. G., Terpstra P., Dijkhuizen L. 1990b; Nucleotide sequences of the genes encoding fructose-bisphosphatase and phosphoribulokinase from Xantbobacter flavus H4-14.. J Gen Microbiol 136:2225–2230
     [Google Scholar]
  27. Meijer W. G., Arnberg A. C., Enequist H. G., Terpstra P., Lidstrom M. E., Dijkhuizen L. 1991; Identification and organization of carbon dioxide fixation genes in Xantbobacter flavus H4-14.. Mol Gen Genet 225:320–330
     [Google Scholar]
  28. Meijer W. G., van den Bergh E. R. E., Smith L. M. 1996; Induction of the gap–pgk operon encoding glyceraldehyde-3-phosphate dehydrogenase and 3-phosphoglycerate kinase of Xantbobacter flavus requires the LysR-type transcriptional activator CbbR.. J Bacteriol 178:881–887
     [Google Scholar]
  29. Paoli G. C., Morgan N. S., Tabita F. R., Shively J. M. 1995; Expression of the cbbLcbbS and cbbM genes and distinct organization of the cbb Calvin cycle structural genes of Rbodo-bacter capsulatus.. Arch Microbiol 164:396–405
     [Google Scholar]
  30. Platt T. 1986; Transcriptional termination and the regulation of gene expression.. Annu Rev Biochem 55:339–372
     [Google Scholar]
  31. Reutz I., Schobert P., Bowien B. 1982; Effect of phosphoglycerate mutase deficiency on heterotrophic and autotrophic carbon metabolism of Alcaligenes eutrophus.. J Bacteriol 151:8–15
     [Google Scholar]
  32. Sambrook J., Fritsch E. F., Maniatis T. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory..
    [Google Scholar]
  33. Schäferjohann J., Yoo J.-G., Kusian B., Bowien B. 1993; The cbb operons of the facultative chemoautotroph Alcaligenes eutrophus encode phosphoglycolate phosphatase.. J Bacteriol l75:7329–7340
     [Google Scholar]
  34. Schäferjohann J., Yoo J.-G., Bowien B. 1995; Analysis of the genes forming the distal parts of the two CO2 fixation operons from Alcaligenes eutrophus.. Arch Microbiol 163:291–299
     [Google Scholar]
  35. Simon R., Priefer U., Pühler A. 1983; A broad host range mobilization system for in vitro engineering: transposon mutagenesis in Gram-negative bacteria.. Biotechnology 1:784–790
     [Google Scholar]
  36. Strecker M., Sickinger E., English R. S., Shively J. M., Bock E. 1994; Calvin cycle genes in Nitrobacter vulgaris T3.. FEMS Microbiol Lett 120:45–50
     [Google Scholar]
  37. Tabita F. R. 1988; Molecular and cellular regulation of autotrophic carbon dioxide fixation in microorganisms.. Microbiol Rev 52:155–189
     [Google Scholar]
  38. Thompson J. D., Higgins D. G., Gibson T. J. 1994; clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-gap penalties and weight matrix choice.. Nucleic Acids Res 22:4673–4680
     [Google Scholar]
  39. Voss H., Schwager C., Wirkner U., Zimmermann J., Erfle H., Hewitt N., Rupp T., Stegemann J., Ansorge W. 1992; New procedure for automated DNA sequencing with multiple internal labeling by fluorescent dUTP.. Methods Mol Cell Biol 3:30–34
     [Google Scholar]
  40. Zimmermann J., Voss H., Schwager C., Stegemann J., Erfle H., Stucky K., Kristensen T., Ansorge W. 1990; A simplified protocol for fast plasmid DNA sequencing.. Nucleic Acids Res 18:1067
     [Google Scholar]
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Xanthobacter flavus employs a single triosephosphate isomerase for heterotrophic and autotrophic metabolism
Microbiology 143, 1925 (1997); https://doi.org/10.1099/00221287-143-6-1925
/content/journal/micro/10.1099/00221287-143-6-1925
/content/journal/micro/10.1099/00221287-143-6-1925
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