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      Involvement of a Novel Genistein-Inducible Multidrug Efflux Pump of Bradyrhizobium japonicum Early in the Interaction with Glycine max (L.) Merr

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          Abstract

          The early molecular dialogue between soybean and the bacterium Bradyrhizobium japonicum is crucial for triggering their symbiotic interaction. Here we found a single large genomic locus that is widely separated from the symbiosis island and was conspicuously induced within minutes after the addition of genistein. This locus (named BjG30) contains genes for the multidrug efflux pump, TetR family transcriptional regulator, and polyhydroxybutyrate (PHB) metabolism. The induction of BjG30 by genistein was competitively inhibited by daidzein, although both genistein and daidzein are soybean-derived inducers of nodulation ( nod) genes. Such a differential expression pattern is also observed in some legume-derived flavonoids, which structurally differ in the hydroxy/deoxy group at the 5-position. In addition, not only did the induction start far in advance of nodW and nodD1 after the addition of genistein, but the levels showed distinct concentration dependence, indicating that the induction pattern of BjG30 is completely different from that of nod genes. The deletion of genes encoding either the multidrug efflux pump or PHB metabolism, especially the former, resulted in defective nodulation performance and nitrogen-fixing capability. Taken together, these results indicate that BjG30, and especially its multidrug efflux pump, may play a key role in the early stage of symbiosis by balancing the dual functions of genistein as both a nod gene inducer and toxicant.

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          Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.

          C Yanisch-Perron, J. Vieira, J Messing (1985)
          Three kinds of improvements have been introduced into the M13-based cloning systems. (1) New Escherichia coli host strains have been constructed for the E. coli bacteriophage M13 and the high-copy-number pUC-plasmid cloning vectors. Mutations introduced into these strains improve cloning of unmodified DNA and of repetitive sequences. A new suppressorless strain facilitates the cloning of selected recombinants. (2) The complete nucleotide sequences of the M13mp and pUC vectors have been compiled from a number of sources, including the sequencing of selected segments. The M13mp18 sequence is revised to include the G-to-T substitution in its gene II at position 6 125 bp (in M13) or 6967 bp in M13mp18. (3) M13 clones suitable for sequencing have been obtained by a new method of generating unidirectional progressive deletions from the polycloning site using exonucleases HI and VII.
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            Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans.

            D Figurski, D Helinski (1979)
            pRK212.2, a derivative of the broad host range plasmid RK2, contains two EcoRI cleavage fragments, A and B, neither of which can replicate by itself in Escherichia coli. Fragment A (41.7 kilobases), but not fragment B (14.4 kilobases), can be cloned by insertion into the unrelated plasmids mini-F and ColE1. Fragment B contains the origin of replication and the ampicillin-resistance determinant of RK2. Transformation of E. coli cells containing the mini-F-fragment A hybrid plasmid with fragment B DNA results in the recircularization and replication of fragment B as a nonmobilizable plasmid (pRK2067) with the copy number and incompatibility properties of RK2. Fragment B cannot be cloned in the absence of fragment A because the latter fragment suppresses a function, specified by fragment B, that results in loss of host cell viability. A small segment (2.4 kilobases) of fragment B that contains the RK2 origin of replication but no longer affects host cell growth in the absence of fragment A had been cloned previously by insertion into a ColE1 plasmid. This hybrid plasmid, designated pRK256, will replicate in E. coli polA mutants only when a fragment A-bearing helper plasmid is present. These results demonstrate that the potentially lethal function specified by fragment B of RK2 is not necessary for replication and that at least one trans-acting function is directly involved in RK2 replication.
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              Efflux-mediated drug resistance in bacteria: an update.

              Xian-Zhi Li, Hiroshi Nikaido (2009)
              Drug efflux pumps play a key role in drug resistance and also serve other functions in bacteria. There has been a growing list of multidrug and drug-specific efflux pumps characterized from bacteria of human, animal, plant and environmental origins. These pumps are mostly encoded on the chromosome, although they can also be plasmid-encoded. A previous article in this journal provided a comprehensive review regarding efflux-mediated drug resistance in bacteria. In the past 5 years, significant progress has been achieved in further understanding of drug resistance-related efflux transporters and this review focuses on the latest studies in this field since 2003. This has been demonstrated in multiple aspects that include but are not limited to: further molecular and biochemical characterization of the known drug efflux pumps and identification of novel drug efflux pumps; structural elucidation of the transport mechanisms of drug transporters; regulatory mechanisms of drug efflux pumps; determining the role of the drug efflux pumps in other functions such as stress responses, virulence and cell communication; and development of efflux pump inhibitors. Overall, the multifaceted implications of drug efflux transporters warrant novel strategies to combat multidrug resistance in bacteria.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Microbes Environ
                Journal ID (iso-abbrev): Microbes Environ
                Title: Microbes and Environments
                Publisher: Japanese Society of Microbial Ecology/The Japanese Society of Soil Microbiology
                ISSN (Print): 1342-6311
                ISSN (Electronic): 1347-4405
                Publication date (Print): December 2013
                Publication date (Electronic): 20 December 2013
                Publication date (Electronic preprint): 25 April 2013
                Volume: 28
                Issue: 4
                Pages: 414-421
                Affiliations
                [1 ]Department of Agricultural and Life Sciences, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Nishi 2–11, Obihiro, Hokkaido, 080–8555, Japan
                [2 ]School of Life Science, Lanzhou University, 222 Tianshui South Rd, Gansu, Lanzhou, 730000, China
                [3 ]Graduate School of Agriculture, Tokyo University of Agriculture and Technology, 3–8–1, Harumi-cho, Fuchu, Tokyo, 183–8538, Japan
                [4 ]Graduate School of Life Science, Tohoku University, 2–1–1, Katahira, Aoba-ku, Sendai, Miyagi, 980–8577, Japan
                [5 ]Faculty of Engineering, Kyoto Sangyo University, Kitaku, Kyoto, 603–8555, Japan
                [6 ]Kazusa DNA Research Institute, 2–6–7, Kazusa-kamatari, Kisarazu, Chiba, 292–0818, Japan
                [7 ]Department of Biological Science, Faculty of Science, Nara Women’s University, Kitauoyanishi-machi, Nara, 630–8506, Japan
                [8 ]Graduate School of Science, Osaka University, 1–1, Machikaneyama, Toyonaka, 560–0043, Osaka, Japan
                [9 ]Department of Life Science, Kagawa University, 2393 Ikenobe, Miki-cho, Kita-gun, Kagawa, 761–0795, Japan
                [10 ]Graduate School of Science and Engineering, Kagoshima University, 1–21–24, Korimoto, Kagoshima, 890–0065, Japan
                Author notes
                [* ]Corresponding author. E-mail: taku@ 123456obihiro.ac.jp ; Tel: +81–155–49–5552; Fax: +81–155–49–5577.
                Article
                Publisher ID: 28_414
                DOI: 10.1264/jsme2.ME13057
                PMC ID: 4070704
                PubMed ID: 24225224
                SO-VID: 0b48855e-9e10-4a70-9312-c380374ac355
                Copyright © Copyright © 2013 by the Japanese Society of Microbial Ecology / the Japanese Society of Soil Microbiology
                History
                Date received : 13 November 2013
                Date accepted : 20 July 2013
                Categories
                Subject: Articles

                Keywords: bradyrhizobium,genistein,multidrug efflux pump,tetr family transcriptional regulator
                Data availability:
                Keywords: bradyrhizobium, genistein, multidrug efflux pump, tetr family transcriptional regulator

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