Analysis of the multimer resolution system encoded by the <i>parCBA</i> operon of broad‐host‐range plasmid RP4

Eberl, Leo; Kristensen, Claus; Givskov, Michael; Grohmann, Elisabeth; Gerlitz, Martin; Schwab, Helmut

doi:10.1111/j.1365-2958.1994.tb01002.x

Cited by 89 publications

(75 citation statements)

References 49 publications

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“…The parDE operon has recently been shown to specify a postsegregational inhibitory system that arrests or kills plasmidless segregants (23,43), and the parCBA operon expresses a site-specific recombinase (ParA) capable of resolving plasmid multimers (10,18,40). However, several lines of evidence indicate that resolution by ParA does not account for the stabilizing effect of the parCBA operon.…”

Section: Discussionmentioning

confidence: 98%

“…One of the known functions of the par region is the multimer resolution activity of ParA (10,18,40). If the instability of pRK2526⌬par results solely from the inability to resolve multimers, then preventing multimer formation by abolishing homologous recombination should suppress the instability (2).…”

Section: Resultsmentioning

confidence: 99%

“…(i) ParA-catalyzed multimer resolution requires a cis-acting site that is located in the parC promoter region (10,40). (ii) An active partition system similar to those of the P1 (1) and F (21) plasmids would also require a specific sequence on the plasmid.…”

Section: Discussionmentioning

confidence: 99%

“…The par region contains five genes (parABCDE) transcribed in two divergent, autoregulated operons (7,9,15,41). The parCBA operon expresses a site-specific recombinase (ParA) (10,18,40) that also functions as a repressor to autoregulate transcription of the operon (7,9). ParA alone can resolve multimeric plasmid forms by interacting with a specific sequence located in the parC promoter region (10).…”

mentioning

confidence: 99%

“…The parCBA operon expresses a site-specific recombinase (ParA) (10,18,40) that also functions as a repressor to autoregulate transcription of the operon (7,9). ParA alone can resolve multimeric plasmid forms by interacting with a specific sequence located in the parC promoter region (10). However, the resolvase activity of ParA is insufficient to explain the stabilization phenotype conferred by the parCBA operon (15,40), and it has been proposed that ParA is part of an active partition apparatus that also involves the ParB and ParC proteins (15).…”

mentioning

confidence: 99%

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Different relative importances of the par operons and the effect of conjugal transfer on the maintenance of intact promiscuous plasmid RK2

et al. 1995

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The par region of the broad-host-range, IncP␣ plasmid RK2 has been implicated as a stability determinant by its ability to enhance the maintenance of mini-RK2 plasmids or heterologous replicons in a growing population of host cells. The region consists of two operons: parCBA, which encodes a multimer resolution system, and parDE, which specifies a postsegregational response mechanism that is toxic to plasmidless segregants. To assess the importance of this region to the stable maintenance of the complete RK2 plasmid in different hosts, we used the vector-mediated excision (VEX) deletion system to specifically remove the entire par region or each operon separately from an otherwise intact RK2 plasmid carrying a lacZ marker. The par region was found to be important to stable maintenance of RK2lac (pRK2526) in Escherichia coli and five other gram-negative hosts (Agrobacterium tumefaciens, Azotobacter vinelandii, Acinetobacter calcoaceticus, Caulobacter crescentus, and Pseudomonas aeruginosa). However, the relative importance of the parCBA and parDE operons varied from host to host. Deletion of parDE had no effect on the maintenance of pRK2526 in A. calcoaceticus, but it severely reduced pRK2526 maintenance in A. vinelandii and resulted in significant instability in the other hosts. Deletion of parCBA did not alter pRK2526 stability in E. coli, A. tumefaciens, or A. vinelandii but severely reduced plasmid maintenance in A. calcoaceticus and P. aeruginosa. In the latter two hosts and C. crescentus, the ⌬parCBA mutant caused a notable reduction in growth rate in the absence of selection for the plasmid, indicating that instability resulting from the absence of parCBA may trigger the postsegregational response mediated by parDE. We also examined the effect of the conjugal transfer system on RK2 maintenance in E. coli. Transfer-defective traJ and traG mutants of pRK2526 were stably maintained in rapidly growing broth cultures. On solid medium, which should be optimal for IncP-mediated conjugation, colonies from cells containing the pRK2526 tra mutants displayed significant numbers of white (Lac ؊ ) sectors on X-Gal (5-bromo-4-chloro-3-indolyl-␤-D-galactopyranoside) plates, whereas sectors appeared rarely in colonies from tra ؉ plasmid-containing cells. Both the traJ and traG mutations further reduced the maintenance of the already unstable ⌬par derivative. Thus, these experiments with defined mutations in an intact RK2 plasmid have revealed (i) that the par region allows RK2 to adapt to the different requirements for stable maintenance in various hosts and (ii) that conjugal transfer can contribute to the maintenance of RK2 in a growing population, particularly under conditions that are favorable to RK2 transfer.The extraordinary success of plasmids in nature relies largely on plasmid-specified functions that promote their dissemination and ensure their persistence in growing populations of bacteria. The highly promiscuous, self-transmissible plasmids of incompatibility group P (IncP␣ and IncP␤) are particularly remarka...

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Section: Discussionmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Different relative importances of the par operons and the effect of conjugal transfer on the maintenance of intact promiscuous plasmid RK2

et al. 1995

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Genetically Engineered Microorganisms for Biodegradation of Recalcitrant Compounds

Duque

Esteve‐Núñez

Michán

et al. 2003

Encyclopedia of Environmental Microbiology

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Key Concepts Hybrid Pathways for Metabolism of Chloroorganic Compounds Modified Pathways for Alkylaromatics Hybrid Pathways for Nitroaromatics Removal of Organomercurials Improved Removal of Pollutants in the Plant Rhizosphere Tracking GEMs in Microcosms and Behavior of Recombinant Microbes

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Genetically Engineered Microorganisms for Biodegradation of Recalcitrant Compounds

Duque¹,

Esteve-nï¿1⁄2ï¿1⁄2ï¿1⁄2ï¿1⁄2ez²,

Michï¿1⁄2ï¿1⁄2n³

et al. 2002

Encyclopedia of Agrochemicals

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The mineralization of organic molecules by microbes is essential for the carbone cycle to operate. The massive mobilization of compounds stored in natural resources or the introduction of xenobiotics into the biosphere produces unidirectional fluxes that result in the persistence of many chemicals in the biosphere, where they are a source of pollution. Molecular biology offers the tools to optimize the biodegradative capacities of microorganisms, accelerate the evolution of new metabolic activities, and construct totally new pathways through the assemblage of catabolic segments from different microbes.

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Analysis of the multimer resolution system encoded by the parCBA operon of broad‐host‐range plasmid RP4

Cited by 89 publications

References 49 publications

Different relative importances of the par operons and the effect of conjugal transfer on the maintenance of intact promiscuous plasmid RK2

Different relative importances of the par operons and the effect of conjugal transfer on the maintenance of intact promiscuous plasmid RK2

Genetically Engineered Microorganisms for Biodegradation of Recalcitrant Compounds

Genetically Engineered Microorganisms for Biodegradation of Recalcitrant Compounds

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