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. 1997 Feb;179(3):592–600. doi: 10.1128/jb.179.3.592-600.1997

Isolation and characterization of a xylose-dependent promoter from Caulobacter crescentus.

A C Meisenzahl 1, L Shapiro 1, U Jenal 1
PMCID: PMC178736  PMID: 9006009

Abstract

An inducible promoter is a useful tool for the controlled expression of a given gene. Accordingly, we identified, cloned, and sequenced a chromosomal locus, xylX, from Caulobacter crescentus which is required for growth on xylose as the sole carbon source and showed that transcription from a single site is dependent on the presence of xylose in the growth medium. P(xylX) promoter activity was determined as a function of the composition of the growth medium both in single copy and on a plasmid using different reporter genes. One hundred micromolar exogenously added xylose was required for maximal induction of P(xylX) in a strain that is unable to metabolize xylose. P(xylX) activity was induced immediately after the addition of xylose and repressed almost completely when xylose was removed from the growth medium. In addition to the strong transcriptional control, the expression of xylX is also regulated on the translational level.

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Selected References

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  1. Amore R., Kötter P., Küster C., Ciriacy M., Hollenberg C. P. Cloning and expression in Saccharomyces cerevisiae of the NAD(P)H-dependent xylose reductase-encoding gene (XYL1) from the xylose-assimilating yeast Pichia stipitis. Gene. 1991 Dec 20;109(1):89–97. doi: 10.1016/0378-1119(91)90592-y. [DOI] [PubMed] [Google Scholar]
  2. Antoine R., Locht C. Isolation and molecular characterization of a novel broad-host-range plasmid from Bordetella bronchiseptica with sequence similarities to plasmids from gram-positive organisms. Mol Microbiol. 1992 Jul;6(13):1785–1799. doi: 10.1111/j.1365-2958.1992.tb01351.x. [DOI] [PubMed] [Google Scholar]
  3. Bor Y. C., Moraes C., Lee S. P., Crosby W. L., Sinskey A. J., Batt C. A. Cloning and sequencing the Lactobacillus brevis gene encoding xylose isomerase. Gene. 1992 May 1;114(1):127–132. doi: 10.1016/0378-1119(92)90718-5. [DOI] [PubMed] [Google Scholar]
  4. Carter P., Bedouelle H., Winter G. Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res. 1985 Jun 25;13(12):4431–4443. doi: 10.1093/nar/13.12.4431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dahl M. K., Degenkolb J., Hillen W. Transcription of the xyl operon is controlled in Bacillus subtilis by tandem overlapping operators spaced by four base-pairs. J Mol Biol. 1994 Oct 28;243(3):413–424. doi: 10.1006/jmbi.1994.1669. [DOI] [PubMed] [Google Scholar]
  6. Dahl M. K., Schmiedel D., Hillen W. Glucose and glucose-6-phosphate interaction with Xyl repressor proteins from Bacillus spp. may contribute to regulation of xylose utilization. J Bacteriol. 1995 Oct;177(19):5467–5472. doi: 10.1128/jb.177.19.5467-5472.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dahms A. S. 3-Deoxy-D-pentulosonic acid aldolase and its role in a new pathway of D-xylose degradation. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1433–1439. doi: 10.1016/0006-291x(74)90358-1. [DOI] [PubMed] [Google Scholar]
  8. David J. D., Wiesmeyer H. Control of xylose metabolism in Escherichia coli. Biochim Biophys Acta. 1970 Mar 24;201(3):497–499. doi: 10.1016/0304-4165(70)90171-6. [DOI] [PubMed] [Google Scholar]
  9. Ely B. Genetics of Caulobacter crescentus. Methods Enzymol. 1991;204:372–384. doi: 10.1016/0076-6879(91)04019-k. [DOI] [PubMed] [Google Scholar]
  10. Evinger M., Agabian N. Envelope-associated nucleoid from Caulobacter crescentus stalked and swarmer cells. J Bacteriol. 1977 Oct;132(1):294–301. doi: 10.1128/jb.132.1.294-301.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gober J. W., Champer R., Reuter S., Shapiro L. Expression of positional information during cell differentiation of Caulobacter. Cell. 1991 Jan 25;64(2):381–391. doi: 10.1016/0092-8674(91)90646-g. [DOI] [PubMed] [Google Scholar]
  12. Gober J. W., Shapiro L. A developmentally regulated Caulobacter flagellar promoter is activated by 3' enhancer and IHF binding elements. Mol Biol Cell. 1992 Aug;3(8):913–926. doi: 10.1091/mbc.3.8.913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hecht G. B., Lane T., Ohta N., Sommer J. M., Newton A. An essential single domain response regulator required for normal cell division and differentiation in Caulobacter crescentus. EMBO J. 1995 Aug 15;14(16):3915–3924. doi: 10.1002/j.1460-2075.1995.tb00063.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hueck C. J., Hillen W. Catabolite repression in Bacillus subtilis: a global regulatory mechanism for the gram-positive bacteria? Mol Microbiol. 1995 Feb;15(3):395–401. doi: 10.1111/j.1365-2958.1995.tb02252.x. [DOI] [PubMed] [Google Scholar]
  15. Hueck C. J., Hillen W., Saier M. H., Jr Analysis of a cis-active sequence mediating catabolite repression in gram-positive bacteria. Res Microbiol. 1994 Sep;145(7):503–518. doi: 10.1016/0923-2508(94)90028-0. [DOI] [PubMed] [Google Scholar]
  16. Jenal U., Shapiro L. Cell cycle-controlled proteolysis of a flagellar motor protein that is asymmetrically distributed in the Caulobacter predivisional cell. EMBO J. 1996 May 15;15(10):2393–2406. [PMC free article] [PubMed] [Google Scholar]
  17. Jenal U., Stephens C., Shapiro L. Regulation of asymmetry and polarity during the Caulobacter cell cycle. Adv Enzymol Relat Areas Mol Biol. 1995;71:1–39. doi: 10.1002/9780470123171.ch1. [DOI] [PubMed] [Google Scholar]
  18. Kurn N., Shapiro L., Agabian N. Effect of carbon source and the role of cyclic adenosine 3',5'-monophosphate on the Caulobacter cell cycle. J Bacteriol. 1977 Sep;131(3):951–959. doi: 10.1128/jb.131.3.951-959.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kötter P., Amore R., Hollenberg C. P., Ciriacy M. Isolation and characterization of the Pichia stipitis xylitol dehydrogenase gene, XYL2, and construction of a xylose-utilizing Saccharomyces cerevisiae transformant. Curr Genet. 1990 Dec;18(6):493–500. doi: 10.1007/BF00327019. [DOI] [PubMed] [Google Scholar]
  20. Lokman B. C., van Santen P., Verdoes J. C., Krüse J., Leer R. J., Posno M., Pouwels P. H. Organization and characterization of three genes involved in D-xylose catabolism in Lactobacillus pentosus. Mol Gen Genet. 1991 Nov;230(1-2):161–169. doi: 10.1007/BF00290664. [DOI] [PubMed] [Google Scholar]
  21. Malakooti J., Wang S. P., Ely B. A consensus promoter sequence for Caulobacter crescentus genes involved in biosynthetic and housekeeping functions. J Bacteriol. 1995 Aug;177(15):4372–4376. doi: 10.1128/jb.177.15.4372-4376.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. POINDEXTER J. S. BIOLOGICAL PROPERTIES AND CLASSIFICATION OF THE CAULOBACTER GROUP. Bacteriol Rev. 1964 Sep;28:231–295. doi: 10.1128/br.28.3.231-295.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Quon K. C., Marczynski G. T., Shapiro L. Cell cycle control by an essential bacterial two-component signal transduction protein. Cell. 1996 Jan 12;84(1):83–93. doi: 10.1016/s0092-8674(00)80995-2. [DOI] [PubMed] [Google Scholar]
  24. Ramakrishnan G., Zhao J. L., Newton A. Multiple structural proteins are required for both transcriptional activation and negative autoregulation of Caulobacter crescentus flagellar genes. J Bacteriol. 1994 Dec;176(24):7587–7600. doi: 10.1128/jb.176.24.7587-7600.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rygus T., Scheler A., Allmansberger R., Hillen W. Molecular cloning, structure, promoters and regulatory elements for transcription of the Bacillus megaterium encoded regulon for xylose utilization. Arch Microbiol. 1991;155(6):535–542. doi: 10.1007/BF00245346. [DOI] [PubMed] [Google Scholar]
  26. Salser W., Gesteland R. F., Bolle A. In vitro synthesis of bacteriophage lysozyme. Nature. 1967 Aug 5;215(5101):588–591. doi: 10.1038/215588a0. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Scheler A., Hillen W. Regulation of xylose utilization in Bacillus licheniformis: Xyl repressor-xyl-operator interaction studied by DNA modification protection and interference. Mol Microbiol. 1994 Aug;13(3):505–512. doi: 10.1111/j.1365-2958.1994.tb00445.x. [DOI] [PubMed] [Google Scholar]
  29. Scheler A., Rygus T., Allmansberger R., Hillen W. Molecular cloning, structure, promoters and regulatory elements for transcription of the Bacillus licheniformis encoded regulon for xylose utilization. Arch Microbiol. 1991;155(6):526–534. doi: 10.1007/BF00245345. [DOI] [PubMed] [Google Scholar]
  30. Schmiedel D., Hillen W. Contributions of XylR CcpA and cre to diauxic growth of Bacillus megaterium and to xylose isomerase expression in the presence of glucose and xylose. Mol Gen Genet. 1996 Feb 25;250(3):259–266. doi: 10.1007/BF02174383. [DOI] [PubMed] [Google Scholar]
  31. Shamanna D. K., Sanderson K. E. Genetics and regulation of D-xylose utilization in Salmonella typhimurium LT2. J Bacteriol. 1979 Jul;139(1):71–79. doi: 10.1128/jb.139.1.71-79.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shapiro L., Agabian-Keshishian N., Hirsch A., Rosen O. M. Effect of dibutyryladenosine 3':5'-cyclic monophosphate on growth and differentiation in Caulobacter crescentus. Proc Natl Acad Sci U S A. 1972 May;69(5):1225–1229. doi: 10.1073/pnas.69.5.1225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sharma S. B., Signer E. R. Temporal and spatial regulation of the symbiotic genes of Rhizobium meliloti in planta revealed by transposon Tn5-gusA. Genes Dev. 1990 Mar;4(3):344–356. doi: 10.1101/gad.4.3.344. [DOI] [PubMed] [Google Scholar]
  34. Sizemore C., Buchner E., Rygus T., Witke C., Götz F., Hillen W. Organization, promoter analysis and transcriptional regulation of the Staphylococcus xylosus xylose utilization operon. Mol Gen Genet. 1991 Jul;227(3):377–384. doi: 10.1007/BF00273926. [DOI] [PubMed] [Google Scholar]
  35. Stephens C., Reisenauer A., Wright R., Shapiro L. A cell cycle-regulated bacterial DNA methyltransferase is essential for viability. Proc Natl Acad Sci U S A. 1996 Feb 6;93(3):1210–1214. doi: 10.1073/pnas.93.3.1210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Van Way S. M., Newton A., Mullin A. H., Mullin D. A. Identification of the promoter and a negative regulatory element, ftr4, that is needed for cell cycle timing of fliF operon expression in Caulobacter crescentus. J Bacteriol. 1993 Jan;175(2):367–376. doi: 10.1128/jb.175.2.367-376.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Vieille C., Hess J. M., Kelly R. M., Zeikus J. G. xylA cloning and sequencing and biochemical characterization of xylose isomerase from Thermotoga neapolitana. Appl Environ Microbiol. 1995 May;61(5):1867–1875. doi: 10.1128/aem.61.5.1867-1875.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wilhelm M., Hollenberg C. P. Selective cloning of Bacillus subtilis xylose isomerase and xylulokinase in Escherichia coli genes by IS5-mediated expression. EMBO J. 1984 Nov;3(11):2555–2560. doi: 10.1002/j.1460-2075.1984.tb02173.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wingrove J. A., Mangan E. K., Gober J. W. Spatial and temporal phosphorylation of a transcriptional activator regulates pole-specific gene expression in Caulobacter. Genes Dev. 1993 Oct;7(10):1979–1992. doi: 10.1101/gad.7.10.1979. [DOI] [PubMed] [Google Scholar]
  40. Winzeler E., Shapiro L. Use of flow cytometry to identify a Caulobacter 4.5 S RNA temperature-sensitive mutant defective in the cell cycle. J Mol Biol. 1995 Aug 18;251(3):346–365. doi: 10.1006/jmbi.1995.0439. [DOI] [PubMed] [Google Scholar]
  41. Wood W. A. Carbohydrate metabolism. Annu Rev Biochem. 1966;35:521–558. doi: 10.1146/annurev.bi.35.070166.002513. [DOI] [PubMed] [Google Scholar]
  42. Zweiger G., Marczynski G., Shapiro L. A Caulobacter DNA methyltransferase that functions only in the predivisional cell. J Mol Biol. 1994 Jan 14;235(2):472–485. doi: 10.1006/jmbi.1994.1007. [DOI] [PubMed] [Google Scholar]

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