Interaction kinetics of the copper-responsive CopY repressor with the cop promoter of Enterococcus hirae

Portmann, Reto; Magnani, David; Stoyanov, Jivko; Schmechel, Ariane; Multhaup, Gerd; Solioz, Marc

doi:10.1007/s00775-004-0536-1

Cited by 44 publications

(51 citation statements)

References 25 publications

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“…Interestingly, there is a perfect cop box upstream of the proposed GGAG ribosome binding site. cop boxes are short, inverted repeats of sequence motif TACAnn TGTA and are the consensus binding motif of CopY-type repressors of firmicutes (16,17). CopY-type repressors are copper inducible and regulate the expression of the copper ATPases in Enterococcus hirae (26) and L. lactis (17) and probably also in related bacteria.…”

Section: Resultsmentioning

confidence: 99%

“…When cytoplasmic copper becomes excessive, the zinc ion of CopY is replaced by two Cu ϩ ions to form Cu ϩ 2 -CopY, which is released from the DNA and allows transcription of the downstream genes to proceed (1,2). CopY-type repressors are ubiquitous in firmicutes and we have previously shown that the purified CopY-type repressors of Enterococcus hirae, Streptococcus mutans, and L. lactis all bind to the same cop box-containing promoters with similar affinity and that binding is abolished by copper (16,17). In agreement with this general concept, purified CopR of L. lactis was found to bind to the cop box present in the promoter region of the lctO gene, and this interaction was abolished by copper.…”

Section: Discussionmentioning

confidence: 99%

“…Induction of the enzyme in turn raised the intracellular pyruvate concentration via the oxidation of lactic acid under use of molecular oxygen. A general copper-regulatory element with the consensus sequence TAC AnnTGTA, also called the cop box (16), was identified in the 5Ј region of the lctO gene and was shown to interact with the CopR regulator. We suggest that the upregulation of LctO by copper serves primarily to defend cells against oxidative stress by decreasing molecular oxygen.…”

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confidence: 99%

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Copper Induction of Lactate Oxidase of Lactococcus lactis : a Novel Metal Stress Response

2007

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Lactococcus lactis IL1403, a lactic acid bacterium widely used for food fermentation, is often exposed to stress conditions. One such condition is exposure to copper, such as in cheese making in copper vats. Copper is an essential micronutrient in prokaryotes and eukaryotes but can be toxic if in excess. Thus, copper homeostatic mechanisms, consisting chiefly of copper transporters and their regulators, have evolved in all organisms to control cytoplasmic copper levels. Using proteomics to identify novel proteins involved in the response of L. lactis IL1403 to copper, cells were exposed to 200 M copper sulfate for 45 min, followed by resolution of the cytoplasmic fraction by two-dimensional gel electrophoresis. One protein strongly induced by copper was LctO, which was shown to be a NAD-independent lactate oxidase. It catalyzed the conversion of lactate to pyruvate in vivo and in vitro. Copper, cadmium, and silver induced LctO, as shown by real-time quantitative PCR. A copper-regulatory element was identified in the 5 region of the lctO gene and shown to interact with the CopR regulator, encoded by the unlinked copRZA operon. Induction of LctO by copper represents a novel copper stress response, and we suggest that it serves in the scavenging of molecular oxygen.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Copper Induction of Lactate Oxidase of Lactococcus lactis : a Novel Metal Stress Response

2007

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“…ORF1 encodes a predicted 227-amino-acid polypeptide with the deduced amino acid sequence of a gene product exhibiting 66 and 63% identity to CopR from P. syringae (33) and to PcoR from E. coli (12), respectively. Using primer extension, the probable transcriptional start of ORF1 was identified 57 bases upstream (G) of the predicted GTG start codon and was associated with a conserved palindrome previously identified as a copper box within copper-responsive promoters (33,34,36) (Fig. 3C).…”

Section: Resultsmentioning

confidence: 99%

Characterization of a Unique Chromosomal Copper Resistance Gene Cluster from X anthomonas campestris pv. vesicatoria

Basım

Minsavage

Stall

et al. 2005

Appl Environ Microbiol

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We characterized the copper resistance genes in strain XvP26 of Xanthomonas campestris pv. vesicatoria, which was originally isolated from a pepper plant in Taiwan. The copper resistance genes were localized to a 7,652-bp region which, based on pulsed-field gel electrophoresis and Southern hybridization, was determined to be located on the chromosome. These genes hybridized only weakly, as determined by Southern analysis, to other copper resistance genes in Xanthomonas and Pseudomonas strains. We identified five open reading frames (ORFs) whose products exhibited high levels of amino acid sequence identity to the products of previously reported copper genes. Mutations in ORF1, ORF3, and ORF4 removed copper resistance, whereas mutations in ORF5 resulted in an intermediate copper resistance phenotype and insertions in ORF2 had no effect on resistance conferred to a copper-sensitive recipient in transconjugant tests. Based on sequence analysis, ORF1 was determined to have high levels of identity with the CopR (66%) and PcoR (63%) genes in Pseudomonas syringae pv. tomato and Escherichia coli, respectively. ORF2 and ORF5 had high levels of identity with the PcoS gene in E. coli and the gene encoding a putative copper-containing oxidoreductase signal peptide protein in Sinorhizobium meliloti, respectively. ORF3 and ORF4 exhibited 23% identity to the gene encoding a cation efflux system membrane protein, CzcC, and 62% identity to the gene encoding a putative copper-containing oxidoreductase protein, respectively. The latter two ORFs were determined to be induced following exposure to low concentrations of copper, while addition of Co, Cd, or Zn resulted in no significant induction. PCR analysis of 51 pepper and 34 tomato copper-resistant X. campestris pv. vesicatoria strains collected from several regions in Taiwan between 1987 and 2000 and nine copper-resistant strains from the United States and South America showed that successful amplification of DNA was obtained only for strain XvP26. The organization of this set of copper resistance genes appears to be uncommon, and the set appears to occur rarely in X. campestris pv. vesicatoria.

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“…Genes involved in copper homeostasis are best characterized in E. hirae (16,(34)(35)(36)(37)(38)(39) but have also been studied in other streptococcal organisms, including S. mutans (14,36) and S. pneumoniae (13,40). In these organisms as well as in other pathogenic bacteria, the genes are located within a cop operon and represent an important virulence mechanism (13,(18)(19)(20)(21).…”

Section: S Pyogenes Clinical Isolates Contain a Cop Operon-like Locusmentioning

confidence: 99%

Copper Tolerance and Characterization of a Copper-Responsive Operon, copYAZ , in an M1T1 Clinical Strain of Streptococcus pyogenes

et al. 2015

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Infection with Streptococcus pyogenes is associated with a breadth of clinical manifestations ranging from mild pharyngitis to severe necrotizing fasciitis. Elevated levels of intracellular copper are highly toxic to this bacterium, and thus, the microbe must tightly regulate the level of this metal ion by one or more mechanisms, which have, to date, not been clearly defined. In this study, we have identified two virulence mechanisms by which S. pyogenes protects itself against copper toxicity. We defined a set of putative genes, copY (for a regulator), copA (for a P1-type ATPase), and copZ (for a copper chaperone), whose expression is regulated by copper. Our results indicate that these genes are highly conserved among a range of clinical S. pyogenes isolates. The copY, copA, and copZ genes are induced by copper and are transcribed as a single unit. Heterologous expression assays revealed that S. pyogenes CopA can confer copper tolerance in a copper-sensitive Escherichia coli mutant by preventing the accumulation of toxic levels of copper, a finding that is consistent with a role for CopA in copper export. Evaluation of the effect of copper stress on S. pyogenes in a planktonic or biofilm state revealed that biofilms may aid in protection during initial exposure to copper. However, copper stress appears to prevent the shift from the planktonic to the biofilm state. Therefore, our results indicate that S. pyogenes may use several virulence mechanisms, including altered gene expression and a transition to and from planktonic and biofilm states, to promote survival during copper stress. IMPORTANCEBacterial pathogens encounter multiple stressors at the host-pathogen interface. This study evaluates a virulence mechanism(s) utilized by S. pyogenes to combat copper at sites of infection. A better understanding of pathogen tolerance to stressors such as copper is necessary to determine how host-pathogen interactions impact bacterial survival during infections. These insights may lead to the identification of novel therapeutic targets that can be used to address antibiotic resistance.

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Interaction kinetics of the copper-responsive CopY repressor with the cop promoter of Enterococcus hirae

Abstract: In Enterococcus hirae,

Cited by 44 publications

References 25 publications

Copper Induction of Lactate Oxidase of Lactococcus lactis : a Novel Metal Stress Response

Copper Induction of Lactate Oxidase of Lactococcus lactis : a Novel Metal Stress Response

Characterization of a Unique Chromosomal Copper Resistance Gene Cluster from X anthomonas campestris pv. vesicatoria

Copper Tolerance and Characterization of a Copper-Responsive Operon, copYAZ , in an M1T1 Clinical Strain of Streptococcus pyogenes

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