Campylobacter jejuni Dps Protein Binds DNA in the Presence of Iron or Hydrogen Peroxide

Huergo, Luciano F.; Rahman, Hossinur; Ibrahimovic, Adis; Day, Christopher J.; Korolik, Victoria

doi:10.1128/jb.00059-13

Cited by 31 publications

(36 citation statements)

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“…These data are in agreement with the results reported for other Dps family proteins (Franceschini et al, 2006;Zhao et al, 2002). However, they are contrary to the results obtained using an Nterminal 10|His-tagged form of CjDps (Huergo et al, 2013). We suspect that these contrasting results were caused by the presence of the His-tag fused with the protein being used in the previous analysis (see below in 'DNA-binding analysis').…”

Section: Effects Of H25g/h37g Substitution On Iron Oxidation Activitycontrasting

confidence: 57%

“…We have also examined the ability of CjDps to interact with DNA by measuring a shift in DNA migration through agarose gel during electrophoresis, similar to that in a previous study where we used the same assay to determine the DNA-binding activity of an engineered N-terminal His-tagged CjDps (Huergo et al, 2013). However, we noted that the untagged CjDps used in the present study showed different responses.…”

Section: Dna-binding Analysismentioning

confidence: 90%

“…Previous data showed that CjDps is able to bind DNA and that this DNA-binding activity is stimulated by Fe 2+ (Huergo et al, 2013). In order to verify if the conserved residues at the Dps FOC participate in iron oxidation and/or could play a role in DNA binding, a variant of CjDps with glycine residues incorporated instead of His25 and His37 was obtained, and confirmed both by DNA sequencing and by MALDI-TOF analysis of the trypsin-digested protein preparations (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The C. jejuni Dps ferroxidase centre Some non-conserved Dps structural motifs have been implicated in DNA-binding activity, such as a lysine-rich N terminus in E. coli Dps (Ceci et al, 2004), a positively charged C terminus in Mycobacterium smegmatis (Roy et al, 2007), or a positively charged protein surface in Helicobacter pylori (Ceci et al, 2007) and C. jejuni Dps (Huergo et al, 2013). In this study, we analysed how the integrity of the FOC affected both the ferroxidase and DNA-binding activities of C. jejuni Dps.…”

Section: Introductionmentioning

confidence: 99%

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Conserved histidine residues at the ferroxidase centre of the Campylobacter jejuni Dps protein are not strictly required for metal binding and oxidation

Sanchuki¹,

Valdameri²,

Moure³

et al. 2016

Microbiology

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Iron is an essential micronutrient for living organisms as it is involved in a broad variety of important biological processes. However, free iron inside the cell could be potentially toxic, generating hydroxyl radicals through the Fenton reaction. Dps (DNA-binding protein from starved cells) belongs to a subfamily of ferritins and can store iron atoms inside the dodecamer. The presence of a ferroxidase centre, composed of highly conserved residues, is a signature of this protein family. In this study, we analysed the role of two conserved histidine residues (H25 and H37) located at the ferroxidase centre of the Campylobacter jejuni Dps protein by replacing them with glycine residues. The C. jejuni H25G/H37G substituted variant showed reduced iron binding and ferroxidase activities in comparison with wt Dps, while DNA-binding activity remained unaffected. We also found that both CjDps wt and CjDps H25G/H37G were able to bind manganese atoms. These results indicate that the H25 and H37 residues at the ferroxidase centre of C. jejuni Dps are not strictly required for metal binding and oxidation.

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Section: Effects Of H25g/h37g Substitution On Iron Oxidation Activitycontrasting

confidence: 57%

Section: Dna-binding Analysismentioning

confidence: 90%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Conserved histidine residues at the ferroxidase centre of the Campylobacter jejuni Dps protein are not strictly required for metal binding and oxidation

Sanchuki¹,

Valdameri²,

Moure³

et al. 2016

Microbiology

Self Cite

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show abstract

“…In C. jejuni, the metalloregulatory proteins Fur and PerR control iron uptake and oxidative stress responses (Holmes et al, 2005;Palyada et al, 2009;van Vliet et al, 1999), with regulation of oxidative stress further complicated by overlapping control circuitry through other regulatory proteins such as CosR and Cj1556 (Dufour et al, 2013;Gundogdu et al, 2011;Hwang et al, 2012;Svensson et al, 2009). Furthermore, DNA-binding by the Dps protein is activated in the presence of iron or hydrogen peroxide, and protects against DNA damage by hydroxyl radicals (Huergo et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

PerR controls oxidative stress defence and aerotolerance but not motility-associated phenotypes of Campylobacter jejuni

et al. 2015

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The foodborne bacterial pathogen Campylobacter jejuni is an obligate microaerophile that is exposed to atmospheric oxygen during transmission through the food chain. Survival under aerobic conditions requires the concerted control of oxidative stress systems, which in C. jejuni are intimately connected with iron metabolism via the PerR and Fur regulatory proteins. Here, we have characterized the roles of C. jejuni PerR in oxidative stress and motility phenotypes, and its regulon at the level of transcription, protein expression and promoter interactions. Insertional inactivation of perR in the C. jejuni reference strains NCTC 11168, 81-176 and 81116 did not result in any growth deficiencies, but strongly increased survival in atmospheric oxygen conditions, and allowed growth around filter discs infused with up to 30 % H2O2 (8.8 M). Expression of catalase, alkyl hydroperoxide reductase, thioredoxin reductase and the Rrc desulforubrerythrin was increased in the perR mutant, and this was mediated at the transcriptional level as shown by electrophoretic mobility shift assays of the katA, ahpC and trxB promoters using purified PerR. Differential RNA-sequencing analysis of a fur perR mutant allowed the identification of eight previously unknown transcription start sites of genes controlled by Fur and/or PerR. Finally, inactivation of perR in C. jejuni did not result in reduced motility, and did not reduce killing of Galleria melonella wax moth larvae. In conclusion, PerR plays an important role in controlling oxidative stress resistance and aerobic survival of C. jejuni, but this role does not extend into control of motility and associated phenotypes

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Controlled modulation of the dynamics of the Deinococcus grandisDps N‐terminal tails by divalent metals

et al. 2023

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DNA‐binding proteins from starved cells (Dps) are small multifunctional nanocages expressed by prokaryotes in acute oxidative stress conditions or during the starvation‐induced stationary phase, as a bacterial defense mechanism. Dps proteins protect bacterial DNA from damage by either direct binding or by removing precursors of reactive oxygen species from solution. The DNA‐binding properties of most Dps proteins studied so far are related to their unordered, flexible, N‐ and C‐terminal extensions. In a previous work, we revealed that the N‐terminal tails of Deinoccocus grandis Dps shift from an extended to a compact conformation depending on the ionic strength of the buffer and detected a novel high‐spin ferrous iron center in the proximal ends of those tails. In this work, we further explore the conformational dynamics of the protein by probing the effect of divalent metals binding to the tail by comparing the metal‐binding properties of the wild‐type protein with a binding site‐impaired D34A variant using size exclusion chromatography, dynamic light scattering, synchrotron radiation circular dichroism, and small‐angle X‐ray scattering. The N‐terminal ferrous species was also characterized by Mössbauer spectroscopy. The results herein presented reveal that the conformation of the N‐terminal tails is altered upon metal binding in a gradual, reversible, and specific manner. These observations may point towards the existence of a regulatory process for the DNA‐binding properties of Dps proteins through metal binding to their N‐ and/or C‐terminal extensions.

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Campylobacter jejuni Dps Protein Binds DNA in the Presence of Iron or Hydrogen Peroxide

Abstract: Iron is an essential cofactor for many enzymes; however, this metal can lead to the formation of reactive oxygen species.

Cited by 31 publications

References 50 publications

Conserved histidine residues at the ferroxidase centre of the Campylobacter jejuni Dps protein are not strictly required for metal binding and oxidation

Conserved histidine residues at the ferroxidase centre of the Campylobacter jejuni Dps protein are not strictly required for metal binding and oxidation

PerR controls oxidative stress defence and aerotolerance but not motility-associated phenotypes of Campylobacter jejuni

Controlled modulation of the dynamics of the Deinococcus grandisDps N‐terminal tails by divalent metals

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