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. 1991 Nov;173(22):7077–7083. doi: 10.1128/jb.173.22.7077-7083.1991

Identification of a novel composite transposable element, Tn5280, carrying chlorobenzene dioxygenase genes of Pseudomonas sp. strain P51.

J R van der Meer 1, A J Zehnder 1, W M de Vos 1
PMCID: PMC209212  PMID: 1657878

Abstract

Analysis of one of the regions of catabolic plasmid pP51 which encode chlorobenzene metabolism of Pseudomonas sp. strain P51 revealed that the tcbA and tcbB genes for chlorobenzene dioxygenase and dehydrogenase are located on a transposable element, Tn5280. Tn5280 showed the features of a composite bacterial transposon with iso-insertion elements (IS1066 and IS1067) at each end of the transposon oriented in an inverted position. When a 12-kb HindIII fragment of pP51 containing Tn5280 was cloned in the suicide donor plasmid pSUP202, marked with a kanamycin resistance gene, and introduced into Pseudomonas putida donor plasmid pSUP202, marked with a kanamycin resistance gene, and introduced into Pseudomonas putida KT2442, Tn5280 was found to transpose into the genome at random and in single copy. The insertion elements IS1066 and IS1067 differed in a single base apir located in the inner inverted repeat and were found to be highly homologous to a class of repetitive elements of Bradyrhizobium japonicum and distantly related to IS630 of Shigella sonnei. The presence of the catabolic genes tcbA and tcbB on Tn5280 suggests a mechanism by which gene clusters can be mobilized as gene cassettes and joined with others to form novel catabolic pathways.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bartels I., Knackmuss H. J., Reineke W. Suicide Inactivation of Catechol 2,3-Dioxygenase from Pseudomonas putida mt-2 by 3-Halocatechols. Appl Environ Microbiol. 1984 Mar;47(3):500–505. doi: 10.1128/aem.47.3.500-505.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berg D. E., Davies J., Allet B., Rochaix J. D. Transposition of R factor genes to bacteriophage lambda. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3628–3632. doi: 10.1073/pnas.72.9.3628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chakrabarty A. M., Friello D. A., Bopp L. H. Transposition of plasmid DNA segments specifying hydrocarbon degradation and their expression in various microorganisms. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3109–3112. doi: 10.1073/pnas.75.7.3109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Don R. H., Weightman A. J., Knackmuss H. J., Timmis K. N. Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134(pJP4). J Bacteriol. 1985 Jan;161(1):85–90. doi: 10.1128/jb.161.1.85-90.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dorn E., Hellwig M., Reineke W., Knackmuss H. J. Isolation and characterization of a 3-chlorobenzoate degrading pseudomonad. Arch Microbiol. 1974;99(1):61–70. doi: 10.1007/BF00696222. [DOI] [PubMed] [Google Scholar]
  7. Franklin F. C., Bagdasarian M., Bagdasarian M. M., Timmis K. N. Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and cloning of genes for the entire regulated aromatic ring meta cleavage pathway. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7458–7462. doi: 10.1073/pnas.78.12.7458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frantz B., Chakrabarty A. M. Organization and nucleotide sequence determination of a gene cluster involved in 3-chlorocatechol degradation. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4460–4464. doi: 10.1073/pnas.84.13.4460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Furukawa K., Miyazaki T. Cloning of a gene cluster encoding biphenyl and chlorobiphenyl degradation in Pseudomonas pseudoalcaligenes. J Bacteriol. 1986 May;166(2):392–398. doi: 10.1128/jb.166.2.392-398.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ghosal D., You I. S. Nucleotide homology and organization of chlorocatechol oxidation genes of plasmids pJP4 and pAC27. Mol Gen Genet. 1988 Jan;211(1):113–120. doi: 10.1007/BF00338401. [DOI] [PubMed] [Google Scholar]
  11. Grindley N. D., Joyce C. M. Genetic and DNA sequence analysis of the kanamycin resistance transposon Tn903. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7176–7180. doi: 10.1073/pnas.77.12.7176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Grindley N. D., Reed R. R. Transpositional recombination in prokaryotes. Annu Rev Biochem. 1985;54:863–896. doi: 10.1146/annurev.bi.54.070185.004243. [DOI] [PubMed] [Google Scholar]
  13. Harayama S., Leppik R. A., Rekik M., Mermod N., Lehrbach P. R., Reineke W., Timmis K. N. Gene order of the TOL catabolic plasmid upper pathway operon and oxidation of both toluene and benzyl alcohol by the xylA product. J Bacteriol. 1986 Aug;167(2):455–461. doi: 10.1128/jb.167.2.455-461.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harayama S., Rekik M., Timmis K. N. Genetic analysis of a relaxed substrate specificity aromatic ring dioxygenase, toluate 1,2-dioxygenase, encoded by TOL plasmid pWW0 of Pseudomonas putida. Mol Gen Genet. 1986 Feb;202(2):226–234. doi: 10.1007/BF00331641. [DOI] [PubMed] [Google Scholar]
  15. Haugland R. A., Sangodkar U. M., Chakrabarty A. M. Repeated sequences including RS1100 from Pseudomonas cepacia AC1100 function as IS elements. Mol Gen Genet. 1990 Jan;220(2):222–228. doi: 10.1007/BF00260485. [DOI] [PubMed] [Google Scholar]
  16. Haugland R. A., Sangodkar U. M., Sferra P. R., Chakrabarty A. M. Cloning and characterization of a chromosomal DNA region required for growth on 2,4,5-T by Pseudomonas cepacia AC1100. Gene. 1991 Apr;100:65–73. doi: 10.1016/0378-1119(91)90351-b. [DOI] [PubMed] [Google Scholar]
  17. Hayase N., Taira K., Furukawa K. Pseudomonas putida KF715 bphABCD operon encoding biphenyl and polychlorinated biphenyl degradation: cloning, analysis, and expression in soil bacteria. J Bacteriol. 1990 Feb;172(2):1160–1164. doi: 10.1128/jb.172.2.1160-1164.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaluza K., Hahn M., Hennecke H. Repeated sequences similar to insertion elements clustered around the nif region of the Rhizobium japonicum genome. J Bacteriol. 1985 May;162(2):535–542. doi: 10.1128/jb.162.2.535-542.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Klecka G. M., Gibson D. T. Inhibition of catechol 2,3-dioxygenase from Pseudomonas putida by 3-chlorocatechol. Appl Environ Microbiol. 1981 May;41(5):1159–1165. doi: 10.1128/aem.41.5.1159-1165.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kleckner N., Chan R. K., Tye B. K., Botstein D. Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J Mol Biol. 1975 Oct 5;97(4):561–575. doi: 10.1016/s0022-2836(75)80059-3. [DOI] [PubMed] [Google Scholar]
  21. Kröckel L., Focht D. D. Construction of chlorobenzene-utilizing recombinants by progenitive manifestation of a rare event. Appl Environ Microbiol. 1987 Oct;53(10):2470–2475. doi: 10.1128/aem.53.10.2470-2475.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Matsutani S., Ohtsubo H., Maeda Y., Ohtsubo E. Isolation and characterization of IS elements repeated in the bacterial chromosome. J Mol Biol. 1987 Aug 5;196(3):445–455. doi: 10.1016/0022-2836(87)90023-4. [DOI] [PubMed] [Google Scholar]
  23. Mondello F. J. Cloning and expression in Escherichia coli of Pseudomonas strain LB400 genes encoding polychlorinated biphenyl degradation. J Bacteriol. 1989 Mar;171(3):1725–1732. doi: 10.1128/jb.171.3.1725-1732.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Osborne D. J., Pickup R. W., Williams P. A. The presence of two complete homologous meta pathway operons on TOL plasmid pWW53. J Gen Microbiol. 1988 Nov;134(11):2965–2975. doi: 10.1099/00221287-134-11-2965. [DOI] [PubMed] [Google Scholar]
  25. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Perkins E. J., Gordon M. P., Caceres O., Lurquin P. F. Organization and sequence analysis of the 2,4-dichlorophenol hydroxylase and dichlorocatechol oxidative operons of plasmid pJP4. J Bacteriol. 1990 May;172(5):2351–2359. doi: 10.1128/jb.172.5.2351-2359.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Reineke W., Knackmuss H. J. Hybrid pathway for chlorobenzoate metabolism in Pseudomonas sp. B13 derivatives. J Bacteriol. 1980 May;142(2):467–473. doi: 10.1128/jb.142.2.467-473.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reineke W., Knackmuss H. J. Microbial degradation of haloaromatics. Annu Rev Microbiol. 1988;42:263–287. doi: 10.1146/annurev.mi.42.100188.001403. [DOI] [PubMed] [Google Scholar]
  29. Sander P., Wittich R. M., Fortnagel P., Wilkes H., Francke W. Degradation of 1,2,4-trichloro- and 1,2,4,5-tetrachlorobenzene by pseudomonas strains. Appl Environ Microbiol. 1991 May;57(5):1430–1440. doi: 10.1128/aem.57.5.1430-1440.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Schraa G., Boone M. L., Jetten M. S., van Neerven A. R., Colberg P. J., Zehnder A. J. Degradation of 1,4-dichlorobenzene by Alcaligenes sp. strain A175. Appl Environ Microbiol. 1986 Dec;52(6):1374–1381. doi: 10.1128/aem.52.6.1374-1381.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Slater J. H., Weightman A. J., Hall B. G. Dehalogenase genes of Pseudomonas putida PP3 on chromosomally located transposable elements. Mol Biol Evol. 1985 Nov;2(6):557–567. doi: 10.1093/oxfordjournals.molbev.a040366. [DOI] [PubMed] [Google Scholar]
  33. Slightom J. L., Durand-Tardif M., Jouanin L., Tepfer D. Nucleotide sequence analysis of TL-DNA of Agrobacterium rhizogenes agropine type plasmid. Identification of open reading frames. J Biol Chem. 1986 Jan 5;261(1):108–121. [PubMed] [Google Scholar]
  34. Tomasek P. H., Frantz B., Sangodkar U. M., Haugland R. A., Chakrabarty A. M. Characterization and nucleotide sequence determination of a repeat element isolated from a 2,4,5-T degrading strain of Pseudomonas cepacia. Gene. 1989;76(2):227–238. doi: 10.1016/0378-1119(89)90163-7. [DOI] [PubMed] [Google Scholar]
  35. Tsuda M., Iino T. Genetic analysis of a transposon carrying toluene degrading genes on a TOL plasmid pWW0. Mol Gen Genet. 1987 Dec;210(2):270–276. doi: 10.1007/BF00325693. [DOI] [PubMed] [Google Scholar]
  36. Tsuda M., Iino T. Identification and characterization of Tn4653, a transposon covering the toluene transposon Tn4651 on TOL plasmid pWW0. Mol Gen Genet. 1988 Jul;213(1):72–77. doi: 10.1007/BF00333400. [DOI] [PubMed] [Google Scholar]
  37. Williams P. A. Genetic interactions between mixed microbial populations. Philos Trans R Soc Lond B Biol Sci. 1982 Jun 11;297(1088):631–639. doi: 10.1098/rstb.1982.0066. [DOI] [PubMed] [Google Scholar]
  38. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  39. Zylstra G. J., McCombie W. R., Gibson D. T., Finette B. A. Toluene degradation by Pseudomonas putida F1: genetic organization of the tod operon. Appl Environ Microbiol. 1988 Jun;54(6):1498–1503. doi: 10.1128/aem.54.6.1498-1503.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. van der Meer J. R., Eggen R. I., Zehnder A. J., de Vos W. M. Sequence analysis of the Pseudomonas sp. strain P51 tcb gene cluster, which encodes metabolism of chlorinated catechols: evidence for specialization of catechol 1,2-dioxygenases for chlorinated substrates. J Bacteriol. 1991 Apr;173(8):2425–2434. doi: 10.1128/jb.173.8.2425-2434.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. van der Meer J. R., Frijters A. C., Leveau J. H., Eggen R. I., Zehnder A. J., de Vos W. M. Characterization of the Pseudomonas sp. strain P51 gene tcbR, a LysR-type transcriptional activator of the tcbCDEF chlorocatechol oxidative operon, and analysis of the regulatory region. J Bacteriol. 1991 Jun;173(12):3700–3708. doi: 10.1128/jb.173.12.3700-3708.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. van der Meer J. R., van Neerven A. R., de Vries E. J., de Vos W. M., Zehnder A. J. Cloning and characterization of plasmid-encoded genes for the degradation of 1,2-dichloro-, 1,4-dichloro-, and 1,2,4-trichlorobenzene of Pseudomonas sp. strain P51. J Bacteriol. 1991 Jan;173(1):6–15. doi: 10.1128/jb.173.1.6-15.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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