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ParI,an orphan ParA family protein from Pseudomonas putida KT2440‐specific genomic island,interferes with the partition system of IncP‐7 plasmids
Authors:Masaki Shintani  Kei Inoue  Tsuguno Terabayashi  Fumiya Sai  Moriya Ohkuma  Hideaki Nojiri  Yuji Nagata  Masataka Tsuda
Affiliation:1. Biotechnology Research Center, The University of Tokyo, , Tokyo, 113‐8657 Japan;2. BioResource Center, Japan Collection of Microorganisms (BRC‐JCM), , Saitama, 351‐0198 Japan;3. Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, , Sendai, 980‐8577 Japan
Abstract:Pseudomonas putida KT2440 is an ideal soil bacterium for expanding the range of degradable compounds via the recruitment of various catabolic plasmids. In the course of our investigation of the host range of IncP‐7 catabolic plasmids pCAR1, pDK1 and pWW53, we found that the IncP‐7 miniplasmids composed of replication and partition loci were exceptionally unstable in KT2440, which is the authentic host of the archetypal IncP‐9 plasmid pWW0. This study identified ParI, a homologue of ParA family of plasmid partitioning proteins encoded on the KT2440‐specific cryptic genomic island, as a negative host factor for the maintenance of IncP‐7 plasmids. The miniplasmids were destabilized by ectopic expression of ParI, and the loss rate correlated with the copy number of ParB binding sites in the centromeric parS region. Mutations in the conserved ATPase domains of ParI abolished destabilization of miniplasmids. Furthermore, ParI destabilized miniplasmid derivatives carrying the partition‐deficient parA mutations but failed to impact the stability of miniplasmid derivatives with parB mutations in the putative arginine finger. Altogether, these results indicate that ParI interferes with the IncP‐7 plasmid partition system. This study extends canonical partition‐mediated incompatibility of plasmids beyond heterogeneous mobile genetic elements, namely incompatibility between plasmid and genomic island.
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