Cloning, expression, purification and characterization of the stress kinase YeaG from Escherichia coli

Tagourti, Jihen; Landoulsi, Ahmed; Richarme, Gilbert

doi:10.1016/j.pep.2008.01.005

Cited by 18 publications

(15 citation statements)

References 13 publications

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“…Using 2D gel electrophoresis, the existence of an autokinase activity of YeaG was confirmed (Tagourti et al 2008). Furthermore, we could detect a 65 kDa protein that was specifically phoshorylated by YeaG, both in extracts from the wild-type strain and in extracts from the mutant that were Fig.…”

Section: Discussionmentioning

confidence: 63%

“…In order to confirm that the autokinase activity attributed to YeaG (Tagourti et al 2008) was not due to a comigrating contaminant, we repeated the autophosphorylation assay and detected protein phosphorylation using 2D-gel electrophoresis. As shown in Fig.…”

Section: Yeag Specifically Phosphorylates a 65-kda Cytoplasmic Proteinmentioning

confidence: 99%

“…Previous attempts to discover YeaG substrates in E. coli extracts using kinase assays followed by SDS-PAGE analysis of phosphoproteins were unsuccessful (Tagourti et al 2008). The low abundance of the putative YeaG substrate(s) and the poor resolution of 1D gels were suspected as reasons for this lack of success.…”

Section: Yeag Specifically Phosphorylates a 65-kda Cytoplasmic Proteinmentioning

confidence: 99%

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Phosphorylation of a 65 kDa cytoplasmic protein by the Escherichia coli YeaG kinase

et al. 2010

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The Escherichia coli YeaG cytoplasmic protein, which is endowed with autokinase and casein kinase activities, has been shown to phosphorylate specifically a 65-kDa cytoplasmic protein. This protein cannot be phosphorylated in an extract from the yeaG mutant, but it can be phosphorylated in an extract from the parental strain or in mutant extract supplemented with purified YeaG. This phosphoprotein displays a pH of 6.8 in 2D gels. Stationary phase induction of YeaG correlates with a significant growth defect of the yeaG mutant at the end of the exponential phase.

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Section: Discussionmentioning

confidence: 63%

Section: Yeag Specifically Phosphorylates a 65-kda Cytoplasmic Proteinmentioning

confidence: 99%

Section: Yeag Specifically Phosphorylates a 65-kda Cytoplasmic Proteinmentioning

confidence: 99%

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Phosphorylation of a 65 kDa cytoplasmic protein by the Escherichia coli YeaG kinase

et al. 2010

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“…We also tried to improve the solubilization of another aggregationprone protein, the bacterial cytoplasmic stress kinase YeaG, which we recently purified and characterized (Tagourti et al, 2008). The gene coding for YeaG was inserted into the expression plasmid pET-21a, under the transcriptional control of the bacteriophage T7 promoter and lac operator (Tagourti et al, 2008).…”

Section: Solubilization Of the Aggregation-prone Kinase Yeag By Overpmentioning

confidence: 99%

“…The gene coding for YeaG was inserted into the expression plasmid pET-21a, under the transcriptional control of the bacteriophage T7 promoter and lac operator (Tagourti et al, 2008). As shown in Fig.…”

Section: Solubilization Of the Aggregation-prone Kinase Yeag By Overpmentioning

confidence: 99%

Membrane docking of an aggregation-prone protein improves its solubilization

Tagourti¹,

Malki²,

Kern³

et al. 2008

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Evolution of the RpoS Regulon: Origin of RpoS and the Conservation of RpoS-Dependent Regulation in Bacteria

Chiang

Schellhorn

2010

J Mol Evol

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The RpoS sigma factor in proteobacteria regulates genes in stationary phase and in response to stress. Although of conserved function, the RpoS regulon may have different gene composition across species due to high genomic diversity and to known environmental conditions that select for RpoS mutants. In this study, the distribution of RpoS homologs in prokaryotes and the differential dependence of regulon members on RpoS for expression in two gamma-proteobacteria (Escherichia coli and Pseudomonas aeruginosa) were examined. Using a maximum-likelihood phylogeny and reciprocal best hits analysis, we show that the RpoS sigma factor is conserved within gamma-, beta-, and delta-proteobacteria. Annotated RpoS of Borrelia and the enteric RpoS are postulated to have separate evolutionary origins. To determine the conservation of RpoS-dependent gene expression across species, reciprocal best hits analysis was used to identify orthologs of the E. coli RpoS regulon in the RpoS regulon of P. aeruginosa. Of the 186 RpoS-dependent genes of E. coli, 50 proteins have an ortholog within the P. aeruginosa genome. Twelve genes of the 50 orthologs are RpoS-dependent in both species, and at least four genes are regulated by RpoS in other gamma-proteobacteria. Despite RpoS conservation in gamma-, beta-, and delta-proteobacteria, RpoS regulon composition is subject to modification between species. Environmental selection for RpoS mutants likely contributes to the evolutionary divergence and specialization of the RpoS regulon within different bacterial genomes.

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Cloning, expression, purification and characterization of the stress kinase YeaG from Escherichia coli

Cited by 18 publications

References 13 publications

Phosphorylation of a 65 kDa cytoplasmic protein by the Escherichia coli YeaG kinase

Phosphorylation of a 65 kDa cytoplasmic protein by the Escherichia coli YeaG kinase

Membrane docking of an aggregation-prone protein improves its solubilization

Evolution of the RpoS Regulon: Origin of RpoS and the Conservation of RpoS-Dependent Regulation in Bacteria

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