title: Effects of three different nucleoid-associated proteins encoded on IncP-7 plasmid pCAR1 on host Pseudomonas putida KT2440.
abstract: Nucleoid-associated proteins (NAPs), which fold bacterial DNA and influence gene transcription, are considered to be global transcriptional regulators of genes on both plasmids and the host chromosome. Incompatibility P-7 group plasmid pCAR1 carries genes encoding three NAPs: H-NS family protein Pmr, NdpA-like protein Pnd, and HU-like protein Phu. In this study, the effects of single or double disruption of pmr, pnd, and phu were assessed in host Pseudomonas putida KT2440. When pmr and pnd or pmr and phu were simultaneously disrupted, both the segregational stability and the structural stability of pCAR1 were markedly decreased, suggesting that Pmr, Pnd, and Phu act as plasmid-stabilizing factors in addition to their established roles in replication and partition systems. The transfer frequency of pCAR1 was significantly decreased in these double mutants. The segregational and structural instability of pCAR1 in the double mutants was recovered by complementation of pmr, whereas no recovery of transfer deficiency was observed. Comprehensive phenotype comparisons showed that the host metabolism of carbon compounds, which was reduced by pCAR1 carriage, was restored by disruption of the NAP gene(s). Transcriptome analyses of mutants indicated that transcription of genes for energy production, conversion, inorganic ion transport, and metabolism were commonly affected; however, how their products altered the phenotypes of mutants was not clear. The findings of this study indicated that Pmr, Pnd, and Phu act synergistically to affect pCAR1 replication, maintenance, and transfer, as well as to alter the host metabolic phenotype.
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors: Chiho
authors: C
authors: Suzuki-Minakuchi
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors: Ryusuke
authors: R
authors: Hirotani
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan Japan Collection of Microorganisms, RIKEN BioResource Center, Ibaraki, Japan Department of Applied Chemistry and Biochemical Engineering, Graduate School of Engineering, Shizuoka University, Hamamatsu, Japan.
authors: Masaki
authors: M
authors: Shintani
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors: Toshiharu
authors: T
authors: Takeda
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, Toyama, Japan.
authors: Yurika
authors: Y
authors: Takahashi
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors: Kazuhiro
authors: K
authors: Matsui
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors: Delyana
authors: D
authors: Vasileva
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan Agricultural Bioinformatics Research Unit, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
authors: Choong-Soo
authors: CS
authors: Yun
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors: Kazunori
authors: K
authors: Okada
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
authors: Hisakazu
authors: H
authors: Yamane
authors: Biotechnology Research Center, The University of Tokyo, Tokyo, Japan Agricultural Bioinformatics Research Unit, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan anojiri@mail.ecc.u-tokyo.ac.jp.
authors: Hideaki
authors: H
authors: Nojiri
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D016958:Pseudomonas putida / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C031908:H-NS protein, bacteria