1887

Abstract

Both of the independently isolated TOL plasmids pWW53 and pDK1 contain multiple regions homologous to the regulatory gene of the archetypal TOL plasmid pWW0. The three homologues on pWW53 vary in the extent of their homology to . is 99% identical to and is located relative to the single copy of in exactly the same way as and on pWW0. The DNA sequence of is 87% identical to the sequence within the coding region but the non-coding DNA upstream is not homologous. There is a frame-shift change at the end of the coding region which causes the C terminus of XylS3 to be extended by an additional 10 amino acids relative to XylS. is anomalous and appears to encode a truncated pseudogene lacking the first 525 bases found in the other genes. Evidence is presented to show that both and act as regulators of pathway operons. Plasmid pDK1 carries two homologues of . is functional and is a hybrid gene: its 5� end and the upstream sequences are highly homologous to both and , whereas its 3� end is identical to . The sequence of is identical to that of the anomalous truncated . Comparison of the organization and the restriction maps of the catabolic operons on pDK1 and pWW53, together with the nucleotide sequences presented here, indicates that the catabolic DNA on pDK1 has derived from a replicon on which the genes are organized similarly to pWW53 and that a genetic rearrangement has taken place involving a reciprocal recombination internal to two of its homologues.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-3-557
1993-03-01
2021-10-27
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/3/mic-139-3-557.html?itemId=/content/journal/micro/10.1099/00221287-139-3-557&mimeType=html&fmt=ahah

References

  1. Assinder S. J., Williams P. A. 1988; Comparison of the meta pathway operons on NAH plasmid pWW60-22 and TOL plasmid pWW53-4 and its evolutionary significance. Journal of General Microbiology 134:2769–2778
    [Google Scholar]
  2. Assinder S. J., Williams P. A. 1990; The TOL plasmids: determinants of the catabolism of toluene and the xylenes. Advances in Microbial Physiology 31:1–69
    [Google Scholar]
  3. Assinder S. J., De Marco P., Sayers J. R., Shaw L. E., Winson M. K., Williams P. A. 1992; Identical resolvases are encoded by Pseudomonas plasmids pWW53 and pDK1. Nucleic Acid Research 20:5476
    [Google Scholar]
  4. Bagdasarian M., Lurz R., Rückert B., Franklin F. C. H., Bagdasarian M. M., Frey J., Timmis K. N. 1981; Specific purpose plasmid cloning vectors. II. Broad host range, high copy number RSF1010-derived vectors and a host: vector system. Gene 16:237–247
    [Google Scholar]
  5. Chatfield L. K., Williams P. A. 1986; Naturally-occurring TOL plasmids isolated from soil carry either two homologous or two nonhomologous catechol 2,3-oxygenase genes. Journal of Bacteriology 168:878–885
    [Google Scholar]
  6. Franklin F. C. H., Bagdasarian M., Bagdasarian M. M., Timmis K. N. 1981; Molecular and functional analysis of the TOL plasmid pWWO from Pseudomonas putida and the cloning of genes for the entire regulated aromatic ring meta cleavage pathway. Proceedings of the National Academy of the Sciences of the United States of America 78:7458–7462
    [Google Scholar]
  7. Harayama S., Rekik M. 1990; The meta cleavage operon of TOL degradative plasmid pWWO comprises 13 genes. Molecular and General Genetics 222:113–120
    [Google Scholar]
  8. Harayama S., Rekik M., Wubbolts M., Rose K., Leppik R. A., Timmis K. N. 1989; Characterization of five genes in the upper pathway operon of TOL plasmid pWWO from Pseudomonas putida and identification of the gene products. Journal of Bacteriology 171:5048–5055
    [Google Scholar]
  9. Henikoff S. 1984; Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
    [Google Scholar]
  10. Inouye S., Nakazawa A., Nakazawa T. 1986; Nucleotide sequence of the regulatory gene xylS on the Pseudomonas putida TOL plasmid and identification of the protein product. Gene 44:235–242
    [Google Scholar]
  11. Jeffrey W. H., Cuskey S. M., Chapman P. J., Resnick S., Olsen R. H. 1992; Characterization of Pseudomonas putida mutants unable to catabolize benzoate: cloning and characterization of Pseudomonas genes involved in benzoate catabolism and isolation of a chromosomal DNA fragment able to substitute for xylS in activation of the TOL-lower pathway promoter. Journal of Bacteriology 174:4986–4996
    [Google Scholar]
  12. Keil H., Keil S., Pickup R. W., Williams P. A. 1985a; Evolutionary conservation of genes coding for meta pathway enzymes within TOL plasmids pWWO and pWW53. Journal of Bacteriology 164:887–895
    [Google Scholar]
  13. Keil H., Lebens M. R., Williams P. A. 1985b; TOL plasmid pWW15 contains two non-homologous, independently-regulated catechol 2,3-oxygenase genes. Journal of Bacteriology 163:248–255
    [Google Scholar]
  14. Keil H., Saint C. M., Williams P. A. 1987a; Gene organization of the first catabolic operon of TOL plasmid pWW53: production of indigo by the xylA gene. Journal of Bacteriology 169:764–770
    [Google Scholar]
  15. Keil H., Keil S., Williams P. A. 1987b; Molecular analysis of regulatory and structural xyl genes of the TOL plasmid pWW53-4. Journal of General Microbiology 133:1149–1158
    [Google Scholar]
  16. Kunz D. A., Chapman P. J. 1981; Isolation and characterization of spontaneously occurring TOL plasmid mutants of Pseudomonas putida HS1. Journal of Bacteriology 146:952–964
    [Google Scholar]
  17. O’Donnell K. J., Williams P. A. 1991; Duplication of both xyl catabolic operons on TOL plasmid pWW15. Journal of General Microbiology 137:2831–2838
    [Google Scholar]
  18. Osborne D. J., Pickup R. W., Williams P. A. 1988; The presence of two homologous meta pathway operons on TOL plasmid pWW53. Journal of General Microbiology 134:2965–2975
    [Google Scholar]
  19. Prentki P., Kirsch A. M. 1982; A modified pBR322 vector with improved properties for the cloning, recovery and sequencing of blunt-ended DNA fragments. Gene 17:189–196
    [Google Scholar]
  20. Ramos J. L., Mermod N., Timmis K. N. 1987; Regulatory circuits controlling transcription of TOL plasmid operon encoding meta cleavage pathway for degradation of alkylbenzoates by Pseudomonas. Molecular Microbiology 1:293–300
    [Google Scholar]
  21. 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]
  22. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 74:5463–5467
    [Google Scholar]
  23. Shaw L. E., Williams P. A. 1988; Physical and functional mapping of two cointegrate plasmids derived from RP4 and TOL plasmid pDK1. Journal of General Microbiology 134:2463–2474
    [Google Scholar]
  24. Spooner R. A., Lindsay K., Franklin F. C. H. 1986; Genetic, functional and sequence analysis of the xylR and xylS regulatory genes of the TOL plasmid pWW0. Journal of General Microbiology 132:1347–1358
    [Google Scholar]
  25. Wheatcroft R., Williams P. A. 1981; Rapid methods for the study of both stable and unstable plasmids in Pseudomonas. Journal of General Microbiology 124:433–437
    [Google Scholar]
  26. Williams P. A., Assinder S. J., Shaw L. E. 1990; Construction of hybrid xylE genes between the two duplicate homologous genes from TOL plasmid pWW53: comparison of the kinetic properties of the gene products. Journal of General Microbiology 136:1583–1589
    [Google Scholar]
  27. Worsey M. J., Williams P. A. 1975; Metabolism of toluene and xylenes by Pseudomonas putida (arvilla) mt-2: evidence for a new function of the TOL plasmid. Journal of Bacteriology 124:7–13
    [Google Scholar]
  28. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-3-557
Loading
/content/journal/micro/10.1099/00221287-139-3-557
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error