Transmembrane Signal Transduction and Osmoregulation in Escherichia coli: II. The Osmotic Sensor, EnvZ, Located in the Isolated Cytoplasmic Membrane Displays Its Phosphorylation and Dephosphorylation Abilities as to the Activator Protein, OmpR1

Tokishita, Shin-ichi; Yamada, H.; Aiba, Hirofumi; Mizuno, Takeshi

doi:10.1093/oxfordjournals.jbchem.a123226

Cited by 54 publications

(47 citation statements)

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“…coli K-12 strain DZ225 (F-, ⌬envZ, lacU169, araD139, rpsL, relA, flgB, thiA), carrying the plasmid-borne multicopy arcB gene, was used to prepare cytoplasmic membranes (Nagasawa et al, 1993). E. coli cells were grown in Luria broth, and the urea-treated cytoplasmic membranes were purified as described previously (Tokishita et al, 1990). The cytoplasmic membranes were used to phosphorylate the Arabidopsis AHP phosphotransmitter proteins in vitro, as described previously (Makino et al, 2000).…”

Section: Escherichia Coli and Related Materialsmentioning

confidence: 99%

“…coli BL21(DE3) cells carrying appropriate plasmids, as described above, were grown in M9-Glc medium containing 0.2% casamino acids and 15% Suc. A cleared cell lysate was obtained using an Amino French pressure cell (FA073; Aminco, SLM Instruments, Urbana, IL) followed by centrifugation at 100,000g for 3 h (Tokishita et al, 1990). In the case of ARR10-RB, the resulting soluble protein samples were applied to a Ni column with the rapid affinity purification pET His-Tag system (Novagen).…”

Section: Purification Of Polypeptidesmentioning

confidence: 99%

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Molecular Structure of the GARP Family of Plant Myb-Related DNA Binding Motifs of the Arabidopsis Response Regulators

et al. 2002

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The B motif is a signature of type-B response regulators (ARRs) involved in His-to-Asp phosphorelay signal transduction systems in Arabidopsis. Homologous motifs occur widely in the GARP family of plant transcription factors. To gain general insight into the structure and function of B motifs (or GARP motifs), we characterized the B motif derived from a representative ARR, ARR10, which led to a number of intriguing findings. First, the B motif of ARR10 (named ARR10-B and extending from Thr-179 to Ser-242) possesses a nuclear localization signal, as indicated by the intracellular localization of a green fluorescent protein-ARR10-B fusion protein in onion epidermal cells. Second, the purified ARR10-B molecule binds specifically in vitro to DNA with the core sequence AGATT. This was demonstrated by several in vitro approaches, including PCR-assisted DNA binding site selection, gel retardation assays, and surface plasmon resonance analysis. Finally, the three-dimensional structure of ARR10-B in solution was determined by NMR spectroscopy, showing that it contains a helix-turn-helix structure. Furthermore, the mode of interaction between ARR10-B and the target DNA was assessed extensively by NMR spectroscopy. Together, these results lead us to propose that the mechanism of DNA recognition by ARR10-B is essentially the same as that of homeodomains. We conclude that the B motif is a multifunctional domain responsible for both nuclear localization and DNA binding and suggest that these insights could be applicable generally to the large GARP family of plant transcription factors.

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Section: Escherichia Coli and Related Materialsmentioning

confidence: 99%

Section: Purification Of Polypeptidesmentioning

confidence: 99%

Molecular Structure of the GARP Family of Plant Myb-Related DNA Binding Motifs of the Arabidopsis Response Regulators

et al. 2002

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“…Urea-treated cytoplasmic membranes containing ArcB were isolated, as previously described (Tokishita et al 1990;Tsuzuki et al 1995). The OmpR protein was puri®ed, as previously described ).…”

Section: In Vitro Phosphorylation Experimentsmentioning

confidence: 99%

Tuning of the porin expression under anaerobic growth conditions by His‐to‐Asp cross‐phosphorelay through both the EnvZ‐osmosensor and ArcB‐anaerosensor in Escherichia coli

et al. 2000

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Background: Widespread bacterial signal transduction circuits are generally referred to as`twocomponent systems' or`histidine (His)-to-aspartate (Asp) phosphorelays.' In Escherichia coli, as many as 30 distinct His-to-Asp phosphorelay signalling pathways operate in response to a wide variety of environmental stimuli, such as medium osmolarity and anaerobiosis. In this regard, it is of interest whether or not some of them together constitute a network of signalling pathways through a physiologically relevant mechanism (often referred to as cross-regulation'). We have addressed this issue, with special reference to the osmo-responsive EnvZ and anaero-responsive ArcB phosphorelay signalling pathways in E. coli.

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“…High levels of OmpR-P favour the expression of OmpC porin. The OmpR-P phosphatase component of the EnvZ protein has recently been shown to be inactivated by high cytoplasmic concentrations of potassium glutamate elicited by changes in external osmotic pressure (Tokishita et al, 1990).…”

Section: -7491 0 1992 Sgmmentioning

confidence: 99%

The regulation of expression of the porin gene ompC by acid pH

Thomas

Booth

1992

Journal of General Microbiology

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The regulation of expression of the porin genes of Escherichiu cofi by acid pH was investigated using reporter gene fusions. The ompC-ZacZ gene fusion was expressed in response to acidification of the external medium. The kinetics of P-galactosidase synthesis under acid-induction differed significantly from those obtained under conditions of osmotic stress. The latter led to rapid induction without a lag, followed by establishment of a rate that was equal to the growth rate; acid-induction was frequently preceded by a short lag period, was relatively slow and did not achieve a rate that was in balance with the growth rate. Further, induction of the ompCgene at acid external pH was dependent upon the presence of glucose as sole carbon source; growth with either glycerol or succinate as sole carbon source reduced induction of ompC at acid pH. Osmotic induction was independent of carbon source. The induction of the ompC gene at acid pH was also reduced by addition of CAMP to the growth medium. The p o M s are known to be subject to catabolite repression and our data are consistent with the exposure to acidic pH resulting in progressive changes in the state of catabolite repression. Acidification of the cytoplasm also provoked a rapid induction of the ompC-ZacZ gene fusion. The kinetics of induction resembled the response to osmotic upshock. This response was independent of the identity of the carbon source supplied for growth. The contribution of changes in cytoplasmic pH to the induction of ompC at acid pH is discussed.

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Transmembrane Signal Transduction and Osmoregulation in Escherichia coli: II. The Osmotic Sensor, EnvZ, Located in the Isolated Cytoplasmic Membrane Displays Its Phosphorylation and Dephosphorylation Abilities as to the Activator Protein, OmpR1

Cited by 54 publications

References 0 publications

Molecular Structure of the GARP Family of Plant Myb-Related DNA Binding Motifs of the Arabidopsis Response Regulators

Molecular Structure of the GARP Family of Plant Myb-Related DNA Binding Motifs of the Arabidopsis Response Regulators

Tuning of the porin expression under anaerobic growth conditions by His‐to‐Asp cross‐phosphorelay through both the EnvZ‐osmosensor and ArcB‐anaerosensor in Escherichia coli

The regulation of expression of the porin gene ompC by acid pH

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