Eri Nishiyama

Publications of Eri Nishiyama

• Pivotal role of anthranilate dioxygenase genes in the adaptation of Burkholderia multivorans ATCC 17616 in soil.

  Authors: Eri Nishiyama, Yoshiyuki Ohtsubo, Yasuhiro Yamamoto, Yuji Nagata, Masataka Tsuda

  FEMS microbiology letters. 02/2012; 330(1):46-55.

  In our recent screen for soil-induced genes, the expression of andA operon (andAcAdAbAa) for anthranilate catabolism in Burkholderia multivorans ATCC 17616 was found to increase dramatically in aIn our recent screen for soil-induced genes, the expression of andA operon (andAcAdAbAa) for anthranilate catabolism in Burkholderia multivorans ATCC 17616 was found to increase dramatically in a soil sample (Nishiyama et al., Environ Microbiol 12: 2539, 2010). The operon was preceded by andR encoding a putative transcriptional regulator for the andA operon. In this study, the andA promoter was induced by tryptophan and anthranilate in an andR-dependent manner. The andA promoter in a deletion mutant lacking tryptophan dioxygenase (one of enzymes for the catabolism of tryptophan to anthranilate) did not respond to tryptophan, indicating that not tryptophan but anthranilate is the effector of AndR. Although both anthranilate and tryptophan were under the detection levels in the soil sample, andA promoter showed higher activity in the soil sample than in a laboratory medium. Such induction required andR and was moderately dependent on the ferric uptake regulator (Fur). The proliferation ability of andAc mutant in the sterile soil was low compared with the co-incubated wild-type cells. These findings suggested that in the soil environment, anthranilate dioxygenase genes are induced by AndR and Fur, and play a pivotal role in the proliferation in the soil environment.

• Identification of Burkholderia multivorans ATCC 17616 genes induced in soil environment by in vivo expression technology.

  Authors: Eri Nishiyama, Yoshiyuki Ohtsubo, Yuji Nagata, Masataka Tsuda

  Environmental microbiology. 04/2010; 12(9):2539-58.

  Burkholderia multivorans ATCC 17616 was originally isolated from a soil sample, and it carries three chromosomes. To identify traits of likely adaptive significance for colonization of soil, an inBurkholderia multivorans ATCC 17616 was originally isolated from a soil sample, and it carries three chromosomes. To identify traits of likely adaptive significance for colonization of soil, an in vivo expression technology system for ATCC 17616 was constructed using the promoterless and tandemly arranged dapB and lacZ genes as the reporters, and this system was applied to identify the genomic loci of ATCC 17616 that were induced in sterilized soil. Our screening of a library consisting of dapB-lacZ-inserted clones resulted in the isolation of 713 clones in which the insertion sites of genome were putatively transcribed in the soil but not in laboratory media. All insertion sites in the genome were determined by high-throughput sequencing using genomic DNA as the templates, and subsequent analysis led to a reliable list of a total of 116 genomic loci as the B. multivorans ATCC 17616 loci induced in a soil environment (mls). These 116 mls carried the genes for energy acquisition from various substances, as well as genes for cell-envelope integrity and the niche adaptation. The distribution of these loci was biased to the second chromosome, suggesting the importance of this replicon as a source of adaptive traits enhancing survival of this organism in natural environments.

• Construction of signature-tagged mutant library in Mesorhizobium loti as a powerful tool for functional genomics.

  Authors: Yoshikazu Shimoda, Hisayuki Mitsui, Hiroko Kamimatsuse, Kiwamu Minamisawa, Eri Nishiyama, Yoshiyuki Ohtsubo, Yuji Nagata, Masataka Tsuda, Sayaka Shinpo, Akiko Watanabe, Mitsuyo Kohara, Manabu Yamada, Yasukazu Nakamura, Satoshi Tabata, Shusei Sato

  DNA research : an international journal for rapid publication of reports on genes and genomes. 10/2008; 15(5):297-308.

  Rhizobia are nitrogen-fixing soil bacteria that establish endosymbiosis with some leguminous plants. The completion of several rhizobial genome sequences provides opportunities for genome-wideRhizobia are nitrogen-fixing soil bacteria that establish endosymbiosis with some leguminous plants. The completion of several rhizobial genome sequences provides opportunities for genome-wide functional studies of the physiological roles of many rhizobial genes. In order to carry out genome-wide phenotypic screenings, we have constructed a large mutant library of the nitrogen-fixing symbiotic bacterium, Mesorhizobium loti, by transposon mutagenesis. Transposon insertion mutants were generated using the signature-tagged mutagenesis (STM) technique and a total of 29,330 independent mutants were obtained. Along with the collection of transposon mutants, we have determined the transposon insertion sites for 7892 clones, and confirmed insertions in 3680 non-redundant M. loti genes (50.5% of the total number of M. loti genes). Transposon insertions were randomly distributed throughout the M. loti genome without any bias toward G+C contents of insertion target sites and transposon plasmids used for the mutagenesis. We also show the utility of STM mutants by examining the specificity of signature tags and test screenings for growth- and nodulation-deficient mutants. This defined mutant library allows for genome-wide forward- and reverse-genetic functional studies of M. loti and will serve as an invaluable resource for researchers to further our understanding of rhizobial biology.