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2 edition of Characterization of the C-domain of the Escherichia coli transcriptional repressor NikR. found in the catalog.

Characterization of the C-domain of the Escherichia coli transcriptional repressor NikR.

Alistair Vincent Michael Dias

Characterization of the C-domain of the Escherichia coli transcriptional repressor NikR.

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Published .
Written in English

About the Edition

NikR is a 15 kDa protein that negatively regulates transcription of the nikABCDE operon, which encodes for an ATP-dependent Ni(II) permease in Escherichia coli. The C-terminal domain of this protein contains the metal binding residues arranged in a novel square planar nickel binding site. In order to further characterize this metal binding activity the C-terminal domain of NikR containing residues 49--133 was cloned, expressed and purified. The metal selectivity of this fragment was then examined using electronic absorption spectroscopy and small molecule competitors.The affinity of the C-domain for metals parallels the Irving-William series of metal complex stabilities, Co(II) < Ni(II) < Cu(II) ≥ Zn(II), indicating that it does not display selectivity for Ni(II) over the other metals investigated. The characteristics of the C-domain are very similar to the previously characterized full-length protein indicating the DNA-binding N-terminal region does not have an effect on high affinity metal binding.

Edition Notes

ContributionsUniversity of Toronto. Dept. of Chemistry.
The Physical Object
Pagination38 leaves.
Number of Pages38
ID Numbers
Open LibraryOL19512328M
ISBN 100612952231

Abstract: Helicobacter pylori is thought to experience little or no competition from other microorganisms in its gastric niche. Only four H. pylori proteins have a perfect match t. The ParG protein ( kDa) is an essential component of the DNA partition complex of multidrug resistance plasmid TP ParG is a dimer in solution, interacts with DNA sequences upstream of the parFG genes and also with the ParF partition protein both in the absence and presence of target DNA. Here, the solution nuclear magnetic resonance structure of ParG is reported. This chapter concentrates on the mechanisms used by Helicobacter pylori to maintain its ion homeostasis, emphasizing metal cations. Maintaining ion homeostasis requires both sensor systems to detect the cytoplasmic ion concentration and effector systems to restore normal cell conditions, or to cope with stress caused by ion imbalance. Iron availability is usually low in most environments, and.

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Characterization of the C-domain of the Escherichia coli transcriptional repressor NikR. by Alistair Vincent Michael Dias Download PDF EPUB FB2

The bacteriophage λ p M promoter is required for maintenance of the λ prophage in Escherichia coli, as it facilitates transcription of the cI Characterization of the C-domain of the Escherichia coli transcriptional repressor NikR.

book, encoding the λ repressor (CI). CI levels are maintained through a transcriptional feedback mechanism whereby CI can serve as an activator or a repressor of p activates p M through cooperative binding to the O R 1 and O R 2 sites within the Cited by: Aiba H.

Autoregulation of the Escherichia coli crp gene: CRP is a transcriptional repressor for its own gene. Cell. Jan; 32 (1)– [] [Google ScholarAldea M, Hernández-Chico C, de la Campa AG, Kushner SR, Vicente M.

Identification, cloning, and expression of bolA, an ftsZ-dependent morphogene of Escherichia by: We have reinvestigated the genetic organization and the transcription regulation of the dsd operon of Escherichia coli.

By combining genetic and biochemical studies, it is demonstrated that the regulatory region of the operon and the gene encoding the specific regulator of D-serine tolerance (dsdC) had been misplaced in previous work on the dsd by:   Recombination of plasmid DNAs and recombination of bacteriophage lambda red mutants in recB recC sbcA Escherichia coli mutants, in which the recE region is expressed, do not require recA.

The recE gene is known to encode exonuclease VIII (exoVIII), which is an ATP-independent exonuclease involved in the RecE pathway of recombination. A 33,molecular-weight (MW) protein was Cited by:   C-Domain and NikR Are Tetramers.

Analytical ultracentrifugation was used to determine the oligomeric state of the purified C-domain and of full-length NikR, both as nickel-bound proteins and as nickel-free proteins (Figures 1C and 1D).In each case, the full-length protein or the C-domain behaved as a tetramer in sedimentation equilibrium experiments performed at protein concentrations Cited by:   1.

Introduction. Escherichia coli OH7 is a highly virulent and toxigenic bacterium that has been involved in a large number of foodborne disease outbreaks (Beuchat, ; Doyle and Erickson, ).This serotype is capable of causing enterohemorrhagic colitis, characterized by bloody diarrhea and occasionally can lead to hemolytic uremic syndrome and kidney failure, especially in.

Escherichia coli group 1 capsules are important virulence determinants, yet little is known about the transcriptional organization or regulation of their biosynthetic (cps.

Transcriptional response of Escherichia coli upon exposure to external copper was studied using DNA microarray and in vivo and in vitro transcription assays. Transcription of three hitherto‐identified copper‐responsive genes, copA (copper efflux transporter), cueO (multicopper oxidase) and cusC (tripartite copper pump component) became maximum at 5 min after addition of copper sulphate.

The transcriptional response of Escherichia coli to elevated levels of metal ions such as Zn(II), Cd(II), Co(II), Ni(II), and Fe(II) has been probed in an effort to understand the mechanisms by which the homeostatic levels of these metal ions are maintained (10, 15, 51, 95, 96).In contrast, very few studies have probed the global response of E.

coli to low levels of metal ions, presumably due. E. Eisenstein, D. BeckettDimerization of the Escherichia coli biotin repressor: corepressor function in protein assembly Biochemistry, 38 (), pp. View Record in Scopus Google Scholar. The X-ray structure of the E.

coli NikR C-domain 6 first identified the high affinity nickel-binding sites, located at the interface between two NikR dimers (see Figure 1).

The Ni 2+ sites are roughly ~40 Å distant from the β sheet in the N-terminus that. The NikR protein is a Ni2+-dependent transcriptional repressor of the nickel-ion uptake system in Escherichia coli, but its mutant protein (NikRm) is able to selectively bind uranyl ions in the.

In the case of E. coli NikR (EcNikR), which is a transcriptional repressor of nickel import (Figure 1B) (), Ni(II) binding induces short-range effects, causing a disorder-to-order transition.

Phage λ infection was investigated for possible production of toxic foreign proteins in Escherichia coli. The target gene transcription was regulated by a T7 promoter, which was initiated under the action of T7 RNA polymerase delivered by infecting phage.

Two types of phage infection were investigated. Transcriptional response of Escherichia coli to extracellular zinc was studied using DNA microarray and S1 mapping assays. Addition of external zinc induced the expression of zinc exporter ZntA and inhibited the expression of zinc importer ZnuC.

In the continuous presence of zinc, ZnuC repression took place at lower zinc concentrations than ZntA induction. Escherichia coli, expressing recombinant green fluorescent protein (GFP), was subjected to dissolved oxygen tension (DOT) oscillations in a two‐compartment system for simulating gradients that can occur in large‐scale were continuously circulated between the anaerobic (0% DOT) and aerobic (10% DOT) vessels of the scale‐down system to mimic an overall.

NikR protein is a Ni 2+ -dependent transcriptional repressor of the nickel-ion uptake system in E. coli [9][10][11] [12]. The tetrameric structure of NikR consists of a core of four metal-binding. -NikR: this family of regulator was initially characterized in E.

coli, is composed of transcriptional regulators described as a dimer of dimers (Chivers and Sauer, ), and repress the. Download Citation | Nickel Regulatory Transcription Factor, NikR | The variety of roles that transition metals fulfill within the cell is staggering, but the concentrations of these metals must be.

The oxyR regulon plays a central role in the defense of Escherichia coli against the endogenous oxidative damage associated with active aerobic growth.

Here we have studied the transcriptional regulation of oxyR in E. coli growing aerobically in rich medium.

Expression of a single-copy oxyR*::lacZ reporter construct varied. Transcriptional responses of Escherichia coli during recovery from inorganic or was also strongly down-regulated under PMA exposure conditions through all times although expression of repressor NikR was Hove-Jensen B.

Ribose catabolism of Escherichia coli: characterization of the rpiB gene encoding ribose phosphate isomerase B and of.

coli NikR repressor binds operator DNA in a nickel-dependent fashion. The pM affinity of NikR for nickel is mediated by its C-terminal 86 residues. Nickel binding induced additional secondary structure, decreased the compactness, and increased the stability of NikR.

Tetramer formation by the C-terminal domain and intact NikR did not require nickel. De Pina K, Desjardin V, Mandrand-Berthelot MA, Giordano G, Wu LF () Isolation and characterization of the nikR gene encoding a nickel-responsive regulator in Escherichia coli.

J Bacteriol – PubMed Google Scholar. Background: MqsR, an endoribonuclease, and MqsA, a transcriptional regulator, form a unique toxin-antitoxin (TA) pair. Results: The high affinity, stable MqsR-MqsA complex is unable to bind DNA. Conclusion: MqsR is the only toxin shown to disrupt the antitoxin-DNA complex, which would promote transcription.

Significance: The MqsR toxin may promote multidrug tolerance in E. coli by disrupting. The Pyrococcus horikoshii OT3 genome contains a gene (PH or nikR) encoding a protein (PhNikR) that shares % amino acid sequence identity with Escherichia coli nickel responsive repressor NikR (EcNikR), including many residues that are functionally important in the E.

coli ortholog. We succeeded in crystallization and structural characterization of PhNikR in the apo. The uptake of nickel in Escherichia coli and other microorganisms is transcriptionally regulated by the NikR repressor or its homologs.

Here we report the. IclR is a repressor for the Escherichia coli aceBAK operon, which encodes isocitrate lyase (aceB), malate synthase (aceA) and isocitrate dehydroge‐nase kinase/phosphorylase (aceK) in the glyoxylate b.

Interactions between DNA-bound transcriptional regulators of the Escherichia coli gal operon. Biochemistry31 (30), DOI: /bia Michael Brenowitz, Amy Pickar, and Elizabeth Jamison.

Stability of a Lac repressor mediated "looped complex". Escherichia coli is one of the organisms of choice for the production of recombinant proteins.

Its use as a cell factory is well-established and it has become the most popular expression platform. For this reason, there are many molecular tools and protocols at hand for the high-level production of heterologous proteins, such as a vast catalog of expression plasmids, a great number of.

Biotin metabolism in prokaryotes is directly linked to transcriptional regulation by the enzyme biotin protein ligase (BPL).

This system is most thoroughly studied and characterized in Escherichia coli. Transcription of the E. coli biotin (bio) operon is directly regulated by the biotin-protein ligase, BirA, the enzyme that covalently attaches.

In Escherichia coli, the sigma factor σ70 directs RNA polymerase to transcribe growth-related genes, while σ38 directs transcription of stress response genes during stationary phase. Two molecules hypothesized to regulate RNA polymerase are the protein Rsd, which binds to σ70, and the non-coding 6S RNA which binds to the RNA polymerase-σ70 holoenzyme.

Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus. F C Lawyer, S Stoffel, R K Saiki, K Myambo, R Drummond and D H Gelfand; Department of Microbial Genetics, Cetus Corporation, Emeryville, California InrS (internal nickel-responsive sensor) is a transcriptional regulator found in cyanobacteria that represses the transcription of the nickel exporter NrsD in the apo form and de-represses expression of the exporter upon Ni(II) binding.

Although a crystal structure of apo-InrS from Synechocystis PCC has been reported, no structure of the protein with metal ions bound is available. Uranium is one of the most important metal resources, and the technology for the recovery of uranyl ions (UO22+) from aqueous solutions is required to ensure a semi-permanent supply of uranium.

The NikR protein is a Ni2+-dependent transcriptional repressor of the nickel-ion uptake system in Escherichia coli, but its mutant protein (NikRm) is able to selectively bind uranyl ions in the.

The in vitro synthesis of Escherichia coli RNA polymerase β and β′ subunits is repressed by either holoenzyme or α2β complex. The level of mRNA synthesized in this system, from the genes encoding ribosomal protein L7/12 (rplL) and RNA polymerase ββ′ subunits (rpoBC) was separately determined by DNA-RNA hybridization using the plasmids, pLOO and pLBC, each containing rplL and rplL.

Stimulation of poly(A) synthesis by Escherichia coli poly(A)polymerase I is correlated with Hfq binding to poly(A) tails. FEBS Journal(2), DOI: /jx. Susan Gottesman. THE SMALL RNA REGULATORS OF ESCHERICHIA COLI: Roles and Mechanisms*. We have investigated the effect of alterations in the structure of the plasmid-borne Escherichia coli tryptophan (trp) coding region and other regions of the same replicon on the level, rate and time of initiation of anthranilate synthetase component I (ASase) synthesis in E.

coli K The maximum level of ASase produced corresponds to 60%–65% of the total cellular proteins. of transcription factors in Escherichia coli have focused on their sequence families and sequence motifs (1,2).

In addition, recent research has elucidated the design principles of the transcriptional regulation network (3–5), including the motifs that recur in the network and their functions. Our approach is based on the determination of the. %B J Biol Chem %V %P %8 12 19 %R / %0 Journal Article %J Inorganic Chemistry %D %T Bimodal Nickel-Binding Site on Escherichia coli [NiFe]-Hydrogenase Metallochaperone HypA %A Lacasse, Michael J.

%A Summers, Kelly L. This chapter describes the characteristics and physiological role of each subfamily, the shared and specialized structural features, and the recognition pattern of target binding sites in detail.

The level of intracellular iron is regulated by various iron-sensing regulators. Intracellular manganese is reported as abundant as zinc in many bacteria, whereas it is about one-tenth of zinc in.

FIGURE 2. Cloning and overexpression of yddV.A, a kb fragment containing the entire yddV structural gene was cloned into pSE In this construct, yddV is under the regulation of a P trc promoter (IPTG-inducible).

B, Coomassie staining of a SDS-PAGE of E. coli cells bearing the pYddV plasmid. A culture of this strain was grown in LB to exponential phase (A = ) and then split in two.Positive control over a wide range of systems in E.

coli is exerted by catabolite repression mediated by the CAP-cAMP system. Negative control in bacteria involves a series of repressor proteins, each specific for a particular pathway.

Several of these systems and the details of their mechanisms of action will be discussed in this chapter.*.The food-borne pathogen Escherichia coli OH7 is commonly exposed to organic acid in processed and preserved foods, allowing adaptation and the development of tolerance to pH levels otherwise lethal.

Since little is known about the molecular basis of adaptation of E. coli to organic acids, we studied K MG and OH7 Sakai during exposure to acetic, lactic, and hydrochloric acid at.