Identification of the Promoter Region and the Transcriptional Regulatory Sequence of the<i>evgAS</i>Operon of<i>Escherichia coli</i>

Tanabe, Hiroyuki; Yamasaki, Katsuhide; Katoh, Akinori; Yoshioka, Setsuko; Utsumi, Ryutaro

doi:10.1271/bbb.62.286

Cited by 11 publications

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“…The 18-bp consensus sequence consists of 5Ј-TT CPyTACA-3Ј and its inverted repeat 5Ј-TGTAPuGAA-3Ј separated by two random bases. Gel mobility shift (36) and DNase I footprinting (13) analyses have shown that EvgA binds to the 27-and 33-bp regions located upstream of evgA/ emrK, respectively. Tanabe et al (36) and Kato et al (13) proposed that the 5Ј-TTCTTA-3Ј sequence and its inverted repeat, 5Ј-TAAGAA-3Ј, separated by a 15-bp spacer sequence, were important for EvgA binding.…”

Section: Discussionmentioning

confidence: 99%

“…Gel mobility shift (36) and DNase I footprinting (13) analyses have shown that EvgA binds to the 27-and 33-bp regions located upstream of evgA/ emrK, respectively. Tanabe et al (36) and Kato et al (13) proposed that the 5Ј-TTCTTA-3Ј sequence and its inverted repeat, 5Ј-TAAGAA-3Ј, separated by a 15-bp spacer sequence, were important for EvgA binding. The inverted repeats that we found are included in both 27-and 33-bp regions that they identified, while the inverted repeats that they proposed are completely included in the 27-bp region but not in the 33-bp regions.…”

Section: Discussionmentioning

confidence: 99%

“…The EvgA overexpression plasmid was constructed as follows. The 884-bp evgA gene with the native promoter and EvgA-binding site, which is important to simulate the evgA promoter (36), was amplified by PCR from MG1655 by using primers 5Ј-TTCCTTAAGCTTCTAAGACTAAACCGTGGCTTTTGCAATAC-3Ј and 5Ј-TTCCTTGAATTCTTAGCCGATTTTGTTACGTTGTGCG-3Ј. These primers contain HindIII and EcoRI sites (underlined) suitable for cloning.…”

Section: Molecular Biology Techniquesmentioning

confidence: 99%

“…The evgAS operon is located adjacent to the emrKY operon, which codes for an efflux pump, and each operon is transcribed in the opposite direction. EvgA binds the intergenic region of evgAS and emrKY and regulates the expression of both operons (13,36). Nishino and Yamaguchi (20) reported that overexpression of EvgA increases multidrug resistance in a drug-hypersusceptible strain which lacks the constitutive multidrug efflux pump genes acrA and acrB (18).…”

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Escherichia coli Gene Expression Responsive to Levels of the Response Regulator EvgA

2002

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To investigate the function of the EvgA response regulator, we compared the genome-wide transcription profile of EvgA-overexpressing and EvgA-lacking Escherichia coli strains by oligonucleotide microarrays. The microarray measurements allowed the identification of at least 37 EvgA-activated genes, including acid resistance-related genes gadABC and hdeAB, efflux pump genes yhiUV and emrK, and 21 genes with unknown function. EvgA overexpression conferred acid resistance to exponentially growing cells. This acid resistance was abolished by deletion of ydeP, ydeO, or yhiE, which was induced by EvgA overexpression. These results suggest that ydeP, ydeO, and yhiE are novel genes related to acid resistance and that EvgA regulates several acid resistance genes. Furthermore, the deletion of yhiE completely abolished acid resistance in stationary-phase cells, suggesting that YhiE plays a critical role in stationary-phase acid resistance. The multidrug resistance in an acrB deletion mutant caused by EvgA overexpression was completely abolished by deletion of yhiUV, while the emrKY deletion had no effect on the increase in resistance by EvgA overexpression. In addition, EvgA overexpression did not confer resistance in a tolC-deficient strain. These results suggest that YhiUV induced by EvgA overexpression is functionally associated with TolC and contributes to multidrug resistance.Bacteria have developed sophisticated signaling systems for adaptive responses to a variety of environments. One of the major mechanisms of signal transduction leading to specific gene expression is known as the two-component system and as its more complex variant, the phosphorelay system (11,24,34,40). A typical two-component system consists of a sensor kinase and its cognate response regulator, which usually functions as a transcriptional factor. The sensor kinase receives an environmental signal, which induces autophosphorylation of a histidine residue. The phosphoryl group on the histidine residue is then transferred to an aspartate residue on the cognate response regulator, resulting in modulation of the expression of its target genes.Genomic sequencing of various microorganisms has revealed the presence of many two-component regulatory systems in all species examined. The functions of many of these putative two-component systems remain unknown. The twocomponent system that consists of response regulator EvgA and sensor kinase EvgS is one such uncharacterized system in Escherichia coli. EvgA and EvgS are highly similar to BvgA and BvgS, respectively, which control the expression of adhesins, toxins, and other virulence factors in Bordetella pertussis (2, 31, 38). The evgAS operon is located adjacent to the emrKY operon, which codes for an efflux pump, and each operon is transcribed in the opposite direction. EvgA binds the intergenic region of evgAS and emrKY and regulates the expression of both operons (13, 36). Nishino and Yamaguchi (20) reported that overexpression of EvgA increases multidrug resistance in a drug-hypersusceptible strain wh...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Molecular Biology Techniquesmentioning

confidence: 99%

mentioning

confidence: 99%

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Escherichia coli Gene Expression Responsive to Levels of the Response Regulator EvgA

2002

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“…3). EvgA is a transcriptional control factor that binds to the repeated sequence (TTCTTA) separated by a 15 bp spacer sequence (EvgA box) upstream of the emrKY and evgA promoters 14,28) (Fig. 3).…”

Section: Evga/evgs Systemmentioning

confidence: 99%

Bacterial Signal Transduction Systems for Environmental Adaptation and Their Inhibitors.

Utsumi

2002

Microb. Environ.

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Bacterial two-component signal transduction systems (TCSs) play a pivotal role in environmental adaptation. In this minireview, two TCSs in Escherichia coli are clarified: the PhoP/PhoQ system, which senses extracellular Mg 2+ and controls the expression of the genes for adaptation to environmental Mg 2+ deprivation (Mg 2+ regulon); and EvgA/EvgS, a signal transduction system that activates expression of the drug efflux pump genes. Furthermore, based on the inhibition of YycF/YycG signal transduction in Bacillus subtilis, a novel strategy for developing a new class of antibacterial agents is described.

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Structure-function relationships in the Bvg and Evg two-component phosphorelay systems

Bantscheff

Perraud

Böck

et al. 2000

International Journal of Medical Microbiology

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Identification of the Promoter Region and the Transcriptional Regulatory Sequence of theevgASOperon ofEscherichia coli

Cited by 11 publications

References 0 publications

Escherichia coli Gene Expression Responsive to Levels of the Response Regulator EvgA

Escherichia coli Gene Expression Responsive to Levels of the Response Regulator EvgA

Bacterial Signal Transduction Systems for Environmental Adaptation and Their Inhibitors.

Structure-function relationships in the Bvg and Evg two-component phosphorelay systems

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