Protein Signatures That Promote Operator Selectivity among Paralog MerR Monovalent Metal Ion Regulators

Humbert, María Victoria; Rasia, Rodolfo M.; Checa, Susana K.; Soncini, Fernando C.

doi:10.1074/jbc.m113.452797

Cited by 21 publications

(26 citation statements)

References 27 publications

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“…The latter regulates gene transcription in response to different environmental signals, including signals from heavy metal ions, organic compounds, and oxidative stress [35]. Approximately 56.5% of the DERs in the Δ VpacspD mutant were down-regulated, of which half were exclusively expressed in the Δ VpacspD mutant (e.g., the two-component response regulator (VpaChn25A_1000) and sigma-E factor negative regulatory protein RseA (VpaChn25_2510)) [36].…”

Section: Resultsmentioning

confidence: 99%

Genomic and transcriptomic analyses reveal distinct biological functions for cold shock proteins (VpaCspA and VpaCspD) in Vibrio parahaemolyticus CHN25 during low-temperature survival

et al. 2017

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Background Vibrio parahaemolyticus causes serious seafood-borne gastroenteritis and death in humans. Raw seafood is often subjected to post-harvest processing and low-temperature storage. To date, very little information is available regarding the biological functions of cold shock proteins (CSPs) in the low-temperature survival of the bacterium. In this study, we determined the complete genome sequence of V. parahaemolyticus CHN25 (serotype: O5:KUT). The two main CSP-encoding genes (VpacspA and VpacspD) were deleted from the bacterial genome, and comparative transcriptomic analysis between the mutant and wild-type strains was performed to dissect the possible molecular mechanisms that underlie low-temperature adaptation by V. parahaemolyticus.ResultsThe 5,443,401-bp V. parahaemolyticus CHN25 genome (45.2% G + C) consisted of two circular chromosomes and three plasmids with 4,724 predicted protein-encoding genes. One dual-gene and two single-gene deletion mutants were generated for VpacspA and VpacspD by homologous recombination. The growth of the ΔVpacspA mutant was strongly inhibited at 10 °C, whereas the VpacspD gene deletion strongly stimulated bacterial growth at this low temperature compared with the wild-type strain. The complementary phenotypes were observed in the reverse mutants (ΔVpacspA-com, and ΔVpacspD-com). The transcriptome data revealed that 12.4% of the expressed genes in V. parahaemolyticus CHN25 were significantly altered in the ΔVpacspA mutant when it was grown at 10 °C. These included genes that were involved in amino acid degradation, secretion systems, sulphur metabolism and glycerophospholipid metabolism along with ATP-binding cassette transporters. However, a low temperature elicited significant expression changes for 10.0% of the genes in the ΔVpacspD mutant, including those involved in the phosphotransferase system and in the metabolism of nitrogen and amino acids. The major metabolic pathways that were altered by the dual-gene deletion mutant (ΔVpacspAD) radically differed from those that were altered by single-gene mutants. Comparison of the transcriptome profiles further revealed numerous differentially expressed genes that were shared among the three mutants and regulators that were specifically, coordinately or antagonistically modulated by VpaCspA and VpaCspD. Our data also revealed several possible molecular coping strategies for low-temperature adaptation by the bacterium.ConclusionsThis study is the first to describe the complete genome sequence of V. parahaemolyticus (serotype: O5:KUT). The gene deletions, complementary insertions, and comparative transcriptomics demonstrate that VpaCspA is a primary CSP in the bacterium, while VpaCspD functions as a growth inhibitor at 10 °C. These results have improved our understanding of the genetic basis for low-temperature survival by the most common seafood-borne pathogen worldwide.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-017-3784-5) contains supplementary material, which is available to a...

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Section: Resultsmentioning

confidence: 99%

Genomic and transcriptomic analyses reveal distinct biological functions for cold shock proteins (VpaCspA and VpaCspD) in Vibrio parahaemolyticus CHN25 during low-temperature survival

et al. 2017

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“…This protein is required for macrophage survival (33), coinciding with an increased copper content in the Salmonella phagosome (15). Despite its function in envelope copper handling, CueP protein expression relies on the cytoplasmic MerR-like regulator CueR (8,34,35); thus, why a major player of periplasmic copper homeostasis is controlled by a cytoplasmic copper sensor was not clear. Here we have demonstrated that CueP expression is controlled not only by CueR, which is activated by the surge in cytoplasmic copper (36), but also, and simultaneously, by CpxR/CpxA, which senses alterations in envelope homeostasis, particularly in the periplasm and the inner membrane (17,18).…”

Section: Discussionmentioning

confidence: 99%

“…EMSAs were performed as described previously (34) using purified CueR and CueR C120S (27,34), CpxR (preincubated with 25 mM acetyl phosphate), and RNA polymerase (Epicentre). DNase I protection assays were done for both DNA strands (34). For KCN treatment, CueR was incubated with KCN to reach a final concentration of 100 or 300 μM in the protein/DNA-binding assay (36).…”

Section: +mentioning

confidence: 99%

Compartment and signal-specific codependence in the transcriptional control of Salmonella periplasmic copper homeostasis

Pezza

Pontel

López

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Copper homeostasis is essential for bacterial pathogen fitness and infection, and has been the focus of a number of recent studies. In Salmonella, envelope protection against copper overload and macrophage survival depends on CueP, a major copper-binding protein in the periplasm. This protein is also required to deliver the metal ion to the Cu/Zn superoxide dismutase SodCII. The Salmonella-specific CueP-coding gene was originally identified as part of the Cue regulon under the transcriptional control of the cytoplasmic copper sensor CueR, but its expression differs from the rest of CueR-regulated genes. Here we show that cueP expression is controlled by the concerted action of CueR, which detects the presence of copper in the cytoplasm, and by CpxR/CpxA, which monitors envelope stress. Copper-activated CueR is necessary for the appropriate spatial arrangement of the −10 and −35 elements of the cueP promoter, and CpxR is essential to recruit the RNA polymerase. The integration of two ancestral sensory systems—CueR, which provides signal specificity, and CpxR/CpxA, which detects stress in the bacterial envelope—restricts the expression of this periplasmic copper resistance protein solely to cells encountering surplus copper that disturbs envelope homeostasis, emulating the role of the CusR/CusS regulatory system present in other enteric bacteria.

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“…These proteins, named CueR in E. coli, detect either Cu(I), Ag(I) or Au(I) in the cytoplasm using two conserved cysteine residues present in a C-terminus loop [54]. They recognize their target operator sequences by an N-terminal helix-turn-helix motif [55]. Structural data available on E. coli CueR revealed that the metal is coordinated in a linear array similar to CopZ-like chaperones [56].…”

Section: Copper Monitoring By Dedicated Sensorsmentioning

confidence: 98%

Bacterial Copper Resistance and Virulence

Pontel

Checa

Soncini

2015

Bacteria-Metal Interactions

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Copper is essential for most organisms. However, it is also toxic even at low levels, especially when its local concentration or intracellular distribution is not properly controlled. Similar to other organisms, bacteria have evolved specific copper homeostasis systems for maintaining a suitable intracellular concentration of this essential metal and at the same time, avoiding its toxic effects. Recent evidence indicates that intracellular copper actively contributes to the host innate immune response against bacterial infections and pathogens have acquired specific mechanisms to deal with this intoxicant. Here, we focus on the different arrays of metal sensing and regulatory systems employed by bacterial pathogens to mount the proper response to counteract the toxic effects of copper allowing survival and replication inside the host.

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Protein Signatures That Promote Operator Selectivity among Paralog MerR Monovalent Metal Ion Regulators

Cited by 21 publications

References 27 publications

Genomic and transcriptomic analyses reveal distinct biological functions for cold shock proteins (VpaCspA and VpaCspD) in Vibrio parahaemolyticus CHN25 during low-temperature survival

Genomic and transcriptomic analyses reveal distinct biological functions for cold shock proteins (VpaCspA and VpaCspD) in Vibrio parahaemolyticus CHN25 during low-temperature survival

Compartment and signal-specific codependence in the transcriptional control of Salmonella periplasmic copper homeostasis

Bacterial Copper Resistance and Virulence

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