Iron inhibits <scp><i>E</i></scp><i>scherichia coli</i> topoisomerase I activity by targeting the first two zinc‐binding sites in the C‐terminal domain

Wu, Wei; Su, Xiaolu; Wang, Xiaobing; Yang, Juanjuan; Zhang, Ting; Wang, Maofeng; Wan, Renhua; Tan, Guoqiang; Lü, Jianxin

doi:10.1002/pro.2542

Cited by 4 publications

(3 citation statements)

References 28 publications

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“…Our previous studies have shown that topoisomerase I (17, 18) and its homolog YrdD (19) are iron and zinc binding proteins, and excess zinc can easily compete for iron binding in the proteins in vivo (17, 19). This suggests that zinc and iron may have similar binding sites in proteins.…”

Section: Introductionmentioning

confidence: 99%

Zinc Toxicity and Iron-Sulfur Cluster Biogenesis in Escherichia coli

Ren

Fan

et al. 2019

Appl Environ Microbiol

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While zinc is an essential trace metal in biology, excess zinc is toxic to organisms. Previous studies have shown that zinc toxicity is associated with disruption of the [4Fe-4S] clusters in various dehydratases in Escherichia coli. Here, we report that the intracellular zinc overload in E. coli cells inhibits iron-sulfur cluster biogenesis without affecting the preassembled iron-sulfur clusters in proteins. Among the housekeeping iron-sulfur cluster assembly proteins encoded by the gene cluster iscSUA-hscBA-fdx-iscX in E. coli cells, the scaffold IscU, the iron chaperone IscA, and ferredoxin have strong zinc binding activity in cells, suggesting that intracellular zinc overload inhibits iron-sulfur cluster biogenesis by binding to the iron-sulfur cluster assembly proteins. Mutations of the conserved cysteine residues to serine in IscA, IscU, or ferredoxin completely abolish the zinc binding activity of the proteins, indicating that zinc can compete with iron or iron-sulfur cluster binding in IscA, IscU, and ferredoxin and block iron-sulfur cluster biogenesis. Furthermore, intracellular zinc overload appears to emulate the slow-growth phenotype of the E. coli mutant cells with deletion of the iron-sulfur cluster assembly proteins IscU, IscA, and ferredoxin. Our results suggest that intracellular zinc overload inhibits iron-sulfur cluster biogenesis by targeting the iron-sulfur cluster assembly proteins IscU, IscA, and ferredoxin in E. coli cells. IMPORTANCE Zinc toxicity has been implicated in causing various human diseases. High concentrations of zinc can also inhibit bacterial cell growth. However, the underlying mechanism has not been fully understood. Here, we report that zinc overload in Escherichia coli cells inhibits iron-sulfur cluster biogenesis by targeting specific iron-sulfur cluster assembly proteins. Because iron-sulfur proteins are involved in diverse physiological processes, the zinc-mediated inhibition of iron-sulfur cluster biogenesis could be largely responsible for the zinc-mediated cytotoxicity. Our finding provides new insights on how intracellular zinc overload may inhibit cellular functions in bacteria.

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Section: Introductionmentioning

confidence: 99%

Zinc Toxicity and Iron-Sulfur Cluster Biogenesis in Escherichia coli

Ren

Fan

et al. 2019

Appl Environ Microbiol

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“…Additionally, the peptide motif Glu-Xaa-Xaa-Glu was found to play a role in the direct binding of ferric iron in several proteins involved in iron transport, sensing, and storage [ 45 ]. The glutamate and histidine residues together form a functional domain for the iron-binding site [ 46 ]. The loop ring between the seventh and eighth transmembrane domains is the second highly variable region (Fig.…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of yellow stripe-like genes in maize suggest their roles in the uptake and transport of zinc and iron

Song,

Li,

et al. 2024

BMC Plant Biol

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Background Yellow Stripe-Like (YSL) proteins are involved in the uptake and transport of metal ions. They play important roles in maintaining the zinc and iron homeostasis in Arabidopsis, rice (Oryza sativa), and barley (Hordeum vulgare). However, proteins in this family have not been fully identified and comprehensively analyzed in maize (Zea mays L.). Results In this study, we identified 19 ZmYSLs in the maize genome and analyzed their structural features. The results of a phylogenetic analysis showed that ZmYSLs are homologous to YSLs of Arabidopsis and rice, and these proteins are divided into four independent branches. Although their exons and introns have structural differences, the motif structure is relatively conserved. Analysis of the cis-regulatory elements in the promoters indicated that ZmYSLs might play a role in response to hypoxia and light. The results of RNA sequencing and quantitative real-time PCR analysis revealed that ZmYSLs are expressed in various tissues and respond differently to zinc and iron deficiency. The subcellular localization of ZmYSLs in the protoplast of maize mesophyll cells showed that they may function in the membrane system. Conclusions This study provided important information for the further functional analysis of ZmYSL, especially in the spatio-temporal expression and adaptation to nutrient deficiency stress. Our findings provided important genes resources for the maize biofortification.

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“…To purify the protein, the culture was pelleted, and bacterial cells were resuspended in Tris buffer (20 mM Tris-HCl, 500 mM NaCl, pH 8.0) before being disrupted by a high-pressure homogenizer (JNBIO). The lysate was then centrifuged at 15,000 × g for 45 min, and the His 6tagged (6 × histidine tag) soluble proteins in the supernatant were purified using a Ni-agarose column (Qiagen co.), followed by a gel filtration column (SuperdexTM 75 10/300GL, GE), as described in Wang et al (2014). The purity of the purified protein was analyzed using SDS/PAGE gel stained by Coomassie brilliant blue.…”

Section: Protein Purificationmentioning

confidence: 99%

A low-cost and eco-friendly recombinant protein expression system using copper-containing industrial wastewater

Zhou,

Xiang,

et al. 2024

Front. Microbiol.

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The development of innovative methods for highly efficient production of recombinant proteins remains a prominent focus of research in the biotechnology field, primarily due to the fact that current commercial protein expression systems rely on expensive chemical inducers, such as isopropyl β-D-thiogalactoside (IPTG). In our study, we designed a novel approach for protein expression by creating a plasmid that responds to copper. This specialized plasmid was engineered through the fusion of a copper-sensing element with an optimized multiple cloning site (MCS) sequence. This MCS sequence can be easily customized by inserting the coding sequences of target recombinant proteins. Once the plasmid was generated, it was introduced into an engineered Escherichia coli strain lacking copA and cueO. With this modified E. coli strain, we demonstrated that the presence of copper ions can efficiently trigger the induction of recombinant protein expression, resulting in the production of active proteins. Most importantly, this expression system can directly utilize copper-containing industrial wastewater as an inducer for protein expression while simultaneously removing copper from the wastewater. Thus, this study provides a low-cost and eco-friendly strategy for the large-scale recombinant protein production. To the best of our knowledge, this is the first report on the induction of recombinant proteins using industrial wastewater.

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Iron inhibits Escherichia coli topoisomerase I activity by targeting the first two zinc‐binding sites in the C‐terminal domain

Cited by 4 publications

References 28 publications

Zinc Toxicity and Iron-Sulfur Cluster Biogenesis in Escherichia coli

Zinc Toxicity and Iron-Sulfur Cluster Biogenesis in Escherichia coli

Identification and characterization of yellow stripe-like genes in maize suggest their roles in the uptake and transport of zinc and iron

A low-cost and eco-friendly recombinant protein expression system using copper-containing industrial wastewater

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