RESUMO
The broad-host-range plasmid RK2 has been a model for studying DNA metabolism in bacteria for many years. It is used as a vector allowing genetic manipulations in numerous bacterial species. The RK2 genome encodes several genes providing the plasmid with diverse functions allowing for its stable maintenance in a variety of bacterial hosts. This review will focus on two processes indispensable for plasmid DNA maintenance. We will summarize recent understanding of the molecular mechanisms contributing to the RK2 DNA replication and partitioning.
Assuntos
Replicação do DNA/fisiologia , Bactérias Gram-Negativas/fisiologia , Fatores R/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Conjugação Genética/genética , Conjugação Genética/fisiologia , Replicação do DNA/genética , Regulação Bacteriana da Expressão Gênica/genética , Regulação Bacteriana da Expressão Gênica/fisiologia , Bactérias Gram-Negativas/genética , Fatores R/genéticaRESUMO
It has been demonstrated that plasmids are not randomly distributed but are located symmetrically in mid-cell, or (1/4), (3/4) positions in bacterial cells. In this work we compared the localization of broad-host-range plasmid RK2 mini-replicons, which lack an active partitioning system, in Escherichia coli and Pseudomonas putida cells. In E. coli the location of the plasmid mini-replicon cluster was at the cell poles. In contrast, in Pseudomonas cells, as a result of the interaction of chromosomally encoded ParB protein with RK2 centromere-like sequences, these mini-derivatives were localized in the proximity of mid-cell, or (1/4), (3/4) positions. The expression of the Pseudomonas parAB genes in E. coli resulted in a positional change in the RK2 mini-derivative to the mid-cell or (1/4), (3/4) positions. Moreover, in a P. putida parAB mutant, both RK2 mini-derivatives and the entire RK2 plasmid exhibited disturbances of subcellular localization. These observations raise the possibility that in certain bacteria chromosomally encoded partitioning machinery could affect subcellular plasmid positioning.