Complete sequence of the IncP‐9 TOL plasmid pWW0 from Pseudomonas putida

Abstract

The TOL plasmid pWW0 (117 kb) is the best studied catabolic plasmid and the archetype of the IncP-9 plasmid incompatibility group from Pseudomonas. It carries the degradative (xyl) genes for toluenes and xylenes within catabolic transposons Tn4651 and Tn4653. Analysis of the complete pWW0 nucleotide sequence revealed 148 putative open reading frames. Of these, 77 showed similarity to published sequences in the available databases predicting functions for: plasmid replication, stable maintenance and transfer; phenotypic determinants; gene regulation and expression; and transposition. All identifiable transposition functions lay within the boundaries of the 70 kb transposon Tn4653, leaving a 46 kb sector containing all the IncP-9 core functions. The replicon and stable inheritance region was very similar to the mini-replicon from IncP-9 antibiotic resistance plasmid pM3, with their Rep proteins forming a novel group of initiation proteins. pWW0 transfer functions exist as two blocks encoding putative DNA processing and mating pair formation genes, with organizational and sequence similarity to IncW plasmids. In addition to the known Tn4651 and IS1246 elements, two additional transposable elements were identified as well as several putative transposition functions, which are probably genetic remnants from previous transposition events. Genes likely to be responsible for known resistance to ultraviolet light and free radicals were identified. Other putative phenotypic functions identified included resistance to mercury and other metal ions, as well as to quaternary ammonium compounds. The complexity and size of pWW0 is largely the result of the mosaic organization of the transposable elements that it carries, rather than the backbone functions of IncP-9 plasmids.

The Behavior and Significance of Degradative Plasmids Belonging to Inc Groups in <i>Pseudomonas</i> within Natural Environments and Microcosms

“…Interestingly, the Dtr systems of the three Inc group plasmids belong to distinct families; IncP-1 (TraI) to MOBP, IncP-7 (TraI) to MOB H , and IncP-9 (TraC) to MOB F (38). The location of the oriT of IncP-1 and IncP-9 plasmids has already been determined experimentally (39,81), whereas that of IncP-7 is still not clear, although the putative oriT region was suggested (56,129). The transcriptional regulation of genes for conjugative transfer on IncP-1 plasmids has been a Substrates of the host of each degradative plasmid: pADP-1, atrazine; pBS265, pBS267, pBS268, ε-caprolactam; pBRC60, pEST4011, pIJB1, pJP4, 2,4-Dichlorophenoxyacetic acid; NAH7, NPL-1, p8C, p15C, pBS240, pBS243, pBS1141, pBS1181, pBS1191, pC1, pDTG1, pNK33, pNK43, pNF143, pOS18, pOS19, pFME4, pFME5, pAK5, pNAH20, pNL4, pNL22, pNL25, pNL29, pNL31, pNL60, pOV17, pSN11, naphthalane, pA81, (halo)aromatic acid; pA8, chlorobenzoate; pSS60, 4-chlorobenzoate; pAC25, pENH91: 3-cholorobenzoate, pPS12-1, tetrachlorobenzene, pTSA, p-toluenesulfonic acid, pDK1, pSVS15, pWW0, pWW53, toluene/xylene, SAL1, salicylate, pCAR1, carbazole.…”

The Crc Global Regulator Inhibits the Pseudomonas putida pWW0 Toluene/Xylene Assimilation Pathway by Repressing the Translation of Regulatory and Structural Genes

“…The CCR imposed on the assimilation of hydrocarbons has been studied with particular emphasis on the pathway for toluene and xylene assimilation specified by the genes carried by the P. putida pWW0 plasmid (8,9). This pathway has become a model system to understand how recalcitrant aromatic compounds are biodegraded in nature.…”

Evidence for self‐association of the alternative sigma factor σ54

“…The P. putida strain KT2440 [20] is a plasmid-free derivative of P. putida mt-2 [32], the original isolate of P. putida harboring the archetypal TOL plasmid pWW0 [33]. E. coli strains DH5a and JM109 were used as host strains for cloning experiments [34].…”