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J Bacteriol. 1992 March; 174(6): 1932-1940

research-article

The dehalogenase gene dehI from Pseudomonas putida PP3 is carried on an unusual mobile genetic element designated DEH.

School of Pure and Applied Biology, University of Wales, Cardiff.

ABSTRACT

As a result of the production of two dehalogenases (DehI and DehII), Pseudomonas putida PP3 utilized halogenated alkanoic acids, such as 2-monochloropropionic acid (2MCPA), as sole sources of carbon and energy. The DehI gene (dehI) was carried on a mobile genetic element (DEH) located on the chromosome of strain PP3. DEH recombined with target plasmid DNAs at high frequencies (e.g. 3.8 x 10(-4) per RP4.5 plasmid transferred). The regulated expression of dehI was detected in P. putida, Pseudomonas aeruginosa, and Escherichia coli strains containing derivative plasmids of RP4.5 and pWW0 recombined with DEH. Movement of DEH from the unstable RP4 derivatives pNJ5000 and pMR5 resulted in the insertion of DEH into the chromosome of RecA+ strains of P. putida but not in RecA+ nor RecA- strains of E. coli. Rescue of DEH from the chromosome of P. putida KT2441 onto plasmid RP4 involved recombination at a frequency (2.7 x 10(-4) per RP4 plasmid transferred) comparable to that observed in strain PP3. The DEH element was not classified as a conventional transposon because it did not move as a discrete DNA fragment: dehI-containing inserts in plasmid DNA targets varied in size between 6 and 13 kb. In addition, DEH exhibited a marked preference for insertion into a specific site on the plasmid pWW0, but its transposition, independent of host recombinational systems, remains to be demonstrated. However, the transposonlike characteristics of DEH included the conservation of restriction endonuclease sites, high-frequency recombination with different target replicons (plasmid and chromosomal DNA), and promiscuous insertion into plasmid RP4-based replicons. Therefore, it is proposed that DEH is an unusual mobile genetic element.

This article has been cited by other articles:

• Weightman, A. J., Topping, A. W., Hill, K. E., Lee, L. L., Sakai, K., Slater, J. H., Thomas, A. W. (2002). Transposition of DEH, a Broad-Host-Range Transposon Flanked by ISPpu12, in Pseudomonas putida Is Associated with Genomic Rearrangements and Dehalogenase Gene Silencing. J. Bacteriol. 184: 6581-6591 [Abstract] [Full Text]  
• Springael, D., Ryngaert, A., Merlin, C., Toussaint, A., Mergeay, M. (2001). Occurrence of Tn4371-Related Mobile Elements and Sequences in (Chloro)biphenyl-Degrading Bacteria. Appl. Environ. Microbiol. 67: 42-50 [Abstract] [Full Text]  
• Hill, K. E., Marchesi, J. R., Weightman, A. J. (1999). Investigation of Two Evolutionarily Unrelated Halocarboxylic Acid Dehalogenase Gene Families. J. Bacteriol. 181: 2535-2547 [Abstract] [Full Text]