Arc and Sfr functions of the Escherichia coli K-12 arcA gene product are genetically and physiologically separable

Silverman, P.; Rother, Stefan; Gaudin, H M

doi:10.1128/jb.173.18.5648-5652.1991

Cited by 25 publications

(14 citation statements)

References 27 publications

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“…Notable modifications include deletions in adhE, ldhA, pflB, mdh, and mutated versions of gltA, lpdA, and arcA. The arcA mutation, resulting in an 8 amino acid insertion in the ArcA protein (Iuchi and Lin, 1988;Silverman et al, 1991) renders the regulator largely inactive, and thus activates the oxidative TCA flux under low O2 conditions (Yim et al, 2011). The gltA and lpdA mutations are the same as described in (Yim et al, 2011).…”

Section: Bacterial Strainmentioning

confidence: 99%

Identification of metabolic engineering targets for the enhancement of 1,4-butanediol production in recombinant E. coli using large-scale kinetic models

Andreozzi

Chakrabarti

Soh

et al. 2016

Metabolic Engineering

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Identification of metabolic engineering targets for the enhancement of 1,4-butanediol production in recombinant E. coli using largescale kinetic models, Metabolic Engineering, http://dx.doi.org/10. 1016/j.ymben.2016.01.009 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting galley proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Entities) to analyze the physiology of recombinant E. coli producing 1,4-butanediol (BDO) and to identify potential strategies for improved production of BDO. The framework allowed us to integrate data across multiple levels and to construct a population of large-scale kinetic models despite the lack of available information about kinetic properties of every enzyme in the metabolic pathways. We analyzed these models and we found that the enzymes that primarily control the fluxes leading to BDO production are part of central glycolysis, the lower branch of tricarboxylic acid (TCA) cycle and the novel BDO production route. Interestingly, among the enzymes between the glucose uptake and the BDO pathway, the enzymes belonging to the lower branch of TCA cycle have been identified as the most important for improving BDO production and yield. We also quantified the effects of changes of the target enzymes on other intracellular states like energy charge, cofactor levels, redox state, cellular growth, and byproduct formation.Independent earlier experiments on this strain confirmed that the computationally 3 obtained conclusions are consistent with the experimentally tested designs, and the findings of the present studies can provide guidance for future work on strain improvement. Overall, these studies demonstrate the potential and effectiveness of ORACLE for the accelerated design of microbial cell factories.

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Section: Bacterial Strainmentioning

confidence: 99%

Identification of metabolic engineering targets for the enhancement of 1,4-butanediol production in recombinant E. coli using large-scale kinetic models

Andreozzi

Chakrabarti

Soh

et al. 2016

Metabolic Engineering

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“…Several host-encoded proteins also influence expression from P Y . The transcriptional activator SfrA (47,53) and integration host factor (49) both influence tra operon expression and F plasmid transfer. Silverman et al (48) observed that cpxA deletion mutants exhibited quasi-wild-type levels of Flac transfer.…”

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confidence: 99%

The Positive Regulator, TraJ, of the Escherichia coli F Plasmid Is Unstable in a cpxA * Background

et al. 2002

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The Cpx (conjugative plasmid expression) stress response of Escherichia coli is induced in response to extracytoplasmic signals generated in the cell envelope, such as misfolded proteins in the periplasm. Detection of stress is mediated by the membrane-bound histidine kinase, CpxA. Signaling of the response regulator CpxR by activated CpxA results in the expression of several factors required for responding to cell envelope stress. CpxA was originally thought to be required for the expression of the positive regulator of the F plasmid transfer (tra) operon, TraJ. It was later determined that constitutive gain-of-function mutations in cpxA led to activation of the Cpx envelope stress response and decreased TraJ expression. In order to determine the nature of the downregulation of TraJ, the level of expression of TraJ, TraM, and TraY, the F-encoded regulatory proteins of the F tra region, was determined both in a cpxA* background and in a wild-type background in which the Cpx stress response was induced by overexpression of the outer membrane lipoprotein, NlpE. Our results suggest that TraJ downregulation is controlled by a posttranscriptional mechanism that operates in the cytoplasm in response to upregulation of the Cpx stress response by both the cpxA* gain-of-function mutation and the overexpression of NlpE.

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confidence: 99%

“…ArcA (or SfrA, for sex factor regulation) is also required for expression of the tra (DNA transfer) genes of the F plasmid. In this function, CpxA, instead of ArcB, might act in conjunction with ArcA (18,30,32,33,38 (21), and in the case of the sensor FrzE of Myxococcus xanthus, an Asp residue(s) has been shown to undergo autophosphorylation (26). In contrast, ArcA is a typical regulator protein, with only the receiver domain containing conserved Asp residues at positions 11 and 54 as potential sites for transphosphorylation by phosphorylated ArcB (ArcB-P) (4,20).…”

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confidence: 99%

Purification and phosphorylation of the Arc regulatory components of Escherichia coli

Iuchi

Lin

1992

J Bacteriol

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In Escherichia coli, a two-component signal transduction system, consisting of the transmembrane sensor protein ArcB and its cognate cytoplasmic regulatory protein ArcA, controls the expression of genes encoding enzymes involved in aerobic respiration. ArcB belongs to a subclass of sensors that have not only a conserved histidine-containing transmitter domain but also a conserved aspartate-containing receiver domain of the regulator family. ' In Escherichia coli, products of the arcA (min 0) and arcB (min 69.5) genes pleiotropically control the expression of numerous target operons, exerting both positive and negative effects. For instance, expression of the structural genes encoding several dehydrogenases of the flavoprotein class; the cytochrome o complex; and members of the tricarboxylic acid cycle, the glyoxylate shunt, and the pathways for fatty acid degradation are anaerobically repressed (15,17). In contrast, expression of the cydAB operon encoding the 02-scavenging cytochrome d is activated under anoxic conditions (6, 16). As a network, these enzymes are responsible for the efficient generation of metabolic energy by aerobic respiration. ArcA (or SfrA, for sex factor regulation) is also required for expression of the tra (DNA transfer) genes of the F plasmid. In this function, CpxA, instead of ArcB, might act in conjunction with ArcA (18,30,32,33,38 (21), and in the case of the sensor FrzE of Myxococcus xanthus, an Asp residue(s) has been shown to undergo autophosphorylation (26). In contrast, ArcA is a typical regulator protein, with only the receiver domain containing conserved Asp residues at positions 11 and 54 as potential sites for transphosphorylation by phosphorylated ArcB (ArcB-P) (4,20). In this study, we constructed plasmids for producing and purifying the two proteins 'ArcB (an ArcB lacking the two transmembrane portions in the N-terminal region) and ArcA for the purpose of testing (i) whether ArcB undergoes autophosphorylation at both His and Asp residues and (ii) whether an Asp residue of ArcA can receive a phosphoryl group from ArcB-P. MATERUILS AND METHODSBacteria, phages, and plasmids. All strains used (XL1-Blue [Stratagene], K38, and ECL594) were derivatives of E. coli K-12. P1 vir phage was used for transductions. To place arcBl in strain K38 with a plasmid (pGP1-2) encoding the T7 polymerase gene (37), the mutant allele in strain ECL594 (15) was cotransduced with a closely linked TnlO (85%). Tetr transductants were selected on LB agar containing tetracycline and purified on the same type of agar. Coinheritance of arcBl was scored for sensitivity to the redox dye toluidine blue o (20). Plasmid pT7-7 was provided by S. Tabor.Growth conditions. For routine cultures, LB medium (1% tryptone, 0.5% yeast extract, 0.5% NaCl) was used. To select Tetr transductants, tetracycline was added at 10 ,ug/ml in LB agar. Ampicillin (100 ,ug/ml) and kanamycin (75 ,ug/ml) were added to LB medium to prevent loss of plasmid vectors bearing a drug resistance marker. To test for dye sensitivity, toluidine bl...

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Arc and Sfr functions of the Escherichia coli K-12 arcA gene product are genetically and physiologically separable

Cited by 25 publications

References 27 publications

Identification of metabolic engineering targets for the enhancement of 1,4-butanediol production in recombinant E. coli using large-scale kinetic models

Identification of metabolic engineering targets for the enhancement of 1,4-butanediol production in recombinant E. coli using large-scale kinetic models

The Positive Regulator, TraJ, of the Escherichia coli F Plasmid Is Unstable in a cpxA * Background

Purification and phosphorylation of the Arc regulatory components of Escherichia coli

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