Transcriptional Analysis of theostA/impGene Involved in Organic Solvent Sensitivity inEscherichia coli

Ohtsu, Iwao; Kakuda, Nobuto; Tsukagoshi, Norihiko; Dokyu, Noriyuki; Takagi, Hiroshi; Wachi, Masaaki; Aono, Rikizo

doi:10.1271/bbb.68.458

Cited by 7 publications

(5 citation statements)

References 9 publications

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“…In contrast, the components involved in LPS transport from the site of biosynthesis in the IM to the bacterial cell surface have only recently been identified (9, 11, 12, 15–17, 47), and the precise functions of the various transport proteins have yet to be resolved. One of the central questions has been whether the process involves a periplasmic scaffold formed by LptA and linking the IM components (LptBFGC) with those in the OM (LptDE).…”

Section: Discussionmentioning

confidence: 99%

Structure and Functional Analysis of LptC, a Conserved Membrane Protein Involved in the Lipopolysaccharide Export Pathway in Escherichia coli*

Tran

Dong

Whitfield

2010

Journal of Biological Chemistry

123

109

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LptC is a conserved bitopic inner membrane protein from Escherichia coli involved in the export of lipopolysaccharide from its site of synthesis in the cytoplasmic membrane to the outer membrane. LptC forms a complex with the ATP-binding cassette transporter, LptBFG, which is thought to facilitate the extraction of lipopolysaccharide from the inner membrane and release it into a translocation pathway that includes the putative periplasmic chaperone LptA. Cysteine modification experiments established that the catalytic domain of LptC is oriented toward the periplasm. The structure of the periplasmic domain is described at a resolution of 2.2-Å from x-ray crystallographic data. The periplasmic domain of LptC consists of a twisted boat structure with two β-sheets in apposition to each other. The β-sheets contain seven and eight antiparallel β-strands, respectively. This structure bears a high degree of resemblance to the crystal structure of LptA. Like LptA, LptC binds lipopolysaccharide in vitro. In vitro, LptA can displace lipopolysaccharide from LptC (but not vice versa), consistent with their locations and their proposed placement in a unidirectional export pathway.

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Section: Discussionmentioning

confidence: 99%

Structure and Functional Analysis of LptC, a Conserved Membrane Protein Involved in the Lipopolysaccharide Export Pathway in Escherichia coli*

Tran

Dong

Whitfield

2010

Journal of Biological Chemistry

123

109

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“…The alteration of organic solvent tolerances of several E. coli mutants by IS integration has been reported. E. coli OST4251 became sensitive to n -hexane because IS 2 and IS 5 became integrated upstream from the imp / ostA gene, which is involved in organic solvent sensitivity (Abe et al 2003 ; Ohtsu et al 2004 ). The hypersensitivity of an E. coli acrB disruptant to organic solvents was suppressed by the integrational activation of the acrEF operon with an IS 1 or IS 2 element (Kobayashi et al 2003 ).…”

Section: Discussionmentioning

confidence: 99%

Contributions of mutations in acrR and marR genes to organic solvent tolerance in Escherichia coli

Watanabe

Doukyu

2012

AMB Express

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The AcrAB-TolC efflux pump is involved in maintaining intrinsic organic solvent tolerance in Escherichia coli. Mutations in regulatory genes such as marR, soxR, and acrR are known to increase the expression level of the AcrAB-TolC pump. To identify these mutations in organic solvent tolerant E. coli, eight cyclohexane-tolerant E. coli JA300 mutants were isolated and examined by DNA sequencing for mutations in marR, soxR, and acrR. Every mutant carried a mutation in either marR or acrR. Among all mutants, strain CH7 carrying a nonsense mutation in marR (named marR109) and an insertion of IS5 in acrR, exhibited the highest organic solvent-tolerance levels. To clarify the involvement of these mutations in improving organic solvent tolerance, they were introduced into the E. coli JA300 chromosome by site-directed mutagenesis using λ red-mediated homologous recombination. Consequently, JA300 mutants carrying acrR::IS5, marR109, or both were constructed and named JA300 acrRIS, JA300 marR, or JA300 acrRIS marR, respectively. The organic solvent tolerance levels of these mutants were increased in the following order: JA300 < JA300 acrRIS < JA300 marR < JA300 acrRIS marR. JA300 acrRIS marR formed colonies on an agar plate overlaid with cyclohexane and p-xylene (6:4 vol/vol mixture). The organic solvent-tolerance level and AcrAB-TolC efflux pump-expression level in JA300 acrRIS marR were similar to those in CH7. Thus, it was shown that the synergistic effects of mutations in only two regulatory genes, acrR and marR, can significantly increase organic solvent tolerance in E. coli.

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“…We also found ZMO_RS01205, OstA encoded organic solvent tolerance protein was up-regulated in mutant strain ZM532 in our omics data (transcriptome and proteome). An earlier studies reported gene ostA is one of the genes contributing to the level of organic solvent tolerance [ 81 , 82 ].…”

Section: Discussionmentioning

confidence: 99%

Molecular mechanism of engineered Zymomonas mobilis to furfural and acetic acid stress

et al. 2023

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Acetic acid and furfural (AF) are two major inhibitors of microorganisms during lignocellulosic ethanol production. In our previous study, we successfully engineered Zymomonas mobilis 532 (ZM532) strain by genome shuffling, but the molecular mechanisms of tolerance to inhibitors were still unknown. Therefore, this study investigated the responses of ZM532 and its wild-type Z. mobilis (ZM4) to AF using multi-omics approaches (transcriptomics, genomics, and label free quantitative proteomics). Based on RNA-Seq data, two differentially expressed genes, ZMO_RS02740 (up-regulated) and ZMO_RS06525 (down-regulated) were knocked out and over-expressed through CRISPR-Cas technology to investigate their roles in AF tolerance. Overall, we identified 1865 and 14 novel DEGs in ZM532 and wild-type ZM4. In contrast, 1532 proteins were identified in ZM532 and wild-type ZM4. Among these, we found 96 important genes in ZM532 involving acid resistance mechanisms and survival rates against stressors. Furthermore, our knockout results demonstrated that growth activity and glucose consumption of mutant strains ZM532∆ZMO_RS02740 and ZM4∆ZMO_RS02740 decreased with increased fermentation time from 42 to 55 h and ethanol production up to 58% in ZM532 than that in ZM532∆ZMO_RS02740. Hence, these findings suggest ZMO_RS02740 as a protective strategy for ZM ethanol production under stressful conditions.

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Transcriptional Analysis of theostA/impGene Involved in Organic Solvent Sensitivity inEscherichia coli

Cited by 7 publications

References 9 publications

Structure and Functional Analysis of LptC, a Conserved Membrane Protein Involved in the Lipopolysaccharide Export Pathway in Escherichia coli*

Structure and Functional Analysis of LptC, a Conserved Membrane Protein Involved in the Lipopolysaccharide Export Pathway in Escherichia coli*

Contributions of mutations in acrR and marR genes to organic solvent tolerance in Escherichia coli

Molecular mechanism of engineered Zymomonas mobilis to furfural and acetic acid stress

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