Volume 14, Issue 11 p. 2946-2959
Research Article

ParI, an orphan ParA family protein from Pseudomonas putida KT2440‐specific genomic island, interferes with the partition system of IncP‐7 plasmids

Masatoshi Miyakoshi

Corresponding Author

Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2‐1‐1 Katahira, Sendai, 980‐8577 Japan

Present address: Institute for Molecular Infection Biology, University of Würzburg, Josef‐Schneider‐Str. 2/Bau D15, 97080 Würzburg, Germany.For correspondence. E‐mail mmiyakoshi@ige.tohoku.ac.jp; Tel. (+81) 22 217 5696; Fax (+81) 22 217 5699.Search for more papers by this author
Masaki Shintani

Biotechnology Research Center, The University of Tokyo, 1‐1‐1 Yayoi, Tokyo, 113‐8657 Japan

BioResource Center, Japan Collection of Microorganisms (BRC‐JCM), Riken, 2‐1 Hirosawa, Wako, Saitama, 351‐0198 Japan

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Kei Inoue

Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2‐1‐1 Katahira, Sendai, 980‐8577 Japan

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Tsuguno Terabayashi

Biotechnology Research Center, The University of Tokyo, 1‐1‐1 Yayoi, Tokyo, 113‐8657 Japan

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Fumiya Sai

Biotechnology Research Center, The University of Tokyo, 1‐1‐1 Yayoi, Tokyo, 113‐8657 Japan

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Moriya Ohkuma

BioResource Center, Japan Collection of Microorganisms (BRC‐JCM), Riken, 2‐1 Hirosawa, Wako, Saitama, 351‐0198 Japan

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Hideaki Nojiri

Biotechnology Research Center, The University of Tokyo, 1‐1‐1 Yayoi, Tokyo, 113‐8657 Japan

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Yuji Nagata

Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2‐1‐1 Katahira, Sendai, 980‐8577 Japan

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Masataka Tsuda

Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2‐1‐1 Katahira, Sendai, 980‐8577 Japan

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First published: 03 August 2012
Citations: 9

Summary

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.