ErpA is important but not essential for the Fe/S cluster biogenesis of Escherichia coli NADH:ubiquinone oxidoreductase (complex I)

Oppermann, Sabrina; Höfflin, Simon; Friedrich, Thorsten

doi:10.1016/j.bbabio.2020.148286

Cited by 3 publications

(3 citation statements)

References 44 publications

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Section: Discussionmentioning

confidence: 93%

“…Iron-sulfur (Fe-S) proteins are crucial for prokaryotic and eukaryotic cell metabolism [21,27,28]. In E. coli, erpA is involved in various metabolic pathways, including respiratory metabolism [8,9,29]. Our transcriptomic analysis of the pa0665 temperaturesensitive mutant revealed changes in gene expression related to the oxidative phosphorylation system, including alterations in the expression of genes involved in the electron transport chain (Figure 6), mainly cytochrome o ubiquinol oxidase and cytochrome c cbb3type oxidase, that participate in the electron transport chain's final steps [30,31].…”

Section: Discussionmentioning

confidence: 99%

“…Comparative protein sequence analysis revealed that Pa0665 shares about 65.5% homologous sequences with ErpA, an A-type iron-sulfur (Fe-S) cluster insertion protein, deemed essential in Escherichia coli [7]; this is crucial for cellular respiration under aerobic conditions, as it contributes to the synthesis of isopentenyl diphosphate (IPP), a precursor for essential electron carriers like ubiquinone and menaquinone [8]. The absence of ErpA markedly diminishes complex I Genes 2024, 15, 590 2 of 14 (NADH: ubiquinone oxidoreductase) content and NADH oxidase activity in the cytoplasmic membrane, significantly impacting the assembly of complex I [9], a testament to its essential role.…”

Section: Introductionmentioning

confidence: 99%

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Genetic Analysis of the ts-Lethal Mutant Δpa0665/pTS-pa0665 Reveals Its Role in Cell Morphology and Oxidative Phosphorylation in Pseudomonas aeruginosa

Zhu,

Zhao,

Yang

2024

Genes

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Pa0665 in Pseudomonas aeruginosa shares homologous sequences with that of the essential A-type iron–sulfur (Fe-S) cluster insertion protein ErpA in Escherichia coli. However, its essentiality in P. aeruginosa and its complementation with E. coli erpA has not been experimentally examined. To fulfill this task, we constructed plasmid-based ts-mutant Δpa0665/pTS-pa0665 using a three-step protocol. The mutant displayed growth defects at 42 °C, which were complemented by expressing ec.erpA. Microscopic observations indicated a petite cell phenotype for Δpa0665/pTS-pa0665 at 42 °C, correlated with the downregulation of the oprG gene. RNA sequencing revealed significant transcriptional changes in genes associated with the oxidative phosphorylation (OXPHOS) system, aligning with reduced ATP levels in Δpa0665/pTS-pa0665 under 42 °C. Additionally, the ts-mutant showed heightened sensitivity to H2O2 at 42 °C. Overall, our study demonstrates the essential role of pa0665 for OXPHOS function and is complemented by ec.erpA. We propose that the plasmid-based ts-allele is useful for genetic analysis of essential genes of interest in P. aeruginosa.

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Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Genetic Analysis of the ts-Lethal Mutant Δpa0665/pTS-pa0665 Reveals Its Role in Cell Morphology and Oxidative Phosphorylation in Pseudomonas aeruginosa

Zhu,

Zhao,

Yang

2024

Genes

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The gene order in the nuo-operon is not essential for the assembly of E. coli complex I

Oppermann

Seng²,

Shweich³

et al. 2022

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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A-Type Carrier Proteins Are Involved in [4Fe-4S] Cluster Insertion into the Radical S -Adenosylmethionine Protein MoaA for the Synthesis of Active Molybdoenzymes

et al. 2021

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Iron sulfur (Fe-S) clusters are important biological cofactors present in proteins with crucial biological functions, from photosynthesis to DNA repair, gene expression and bioenergetic processes. For the insertion of Fe-S clusters into proteins, A-type carrier proteins have been identified. So far, three of them were characterized in detail in Escherichia coli, namely IscA, SufA and ErpA, which were shown to partially replace each other in their roles in [4Fe-4S] cluster insertion into specific target proteins. To further expand the knowledge of [4Fe-4S] cluster insertion into proteins, we analyzed the complex Fe-S cluster dependent network for the synthesis of the molybdenum cofactor (Moco) and the expression of genes encoding nitrate reductase in E. coli. Our studies include the identification of the A-type carrier proteins ErpA and IscA involved in [4Fe-4S] cluster insertion into the S-adenosyl-methionine dependent radical SAM protein MoaA. We show that ErpA and IscA can partially replace each other in their role to provide [4Fe-4S] clusters for MoaA. Since most genes expressing molybdoenzymes are regulated by the transcriptional regulator for fumarate and nitrate reduction (FNR) under anaerobic conditions, we also identified the proteins that are crucial to obtain an active FNR under conditions of nitrate respiration. We show that ErpA is essential for the FNR-dependent expression of the narGHJI operon, a role that cannot be compensated by IscA under the growth conditions tested. SufA does not have a role in Fe-S cluster insertion into MoaA or FNR under anaerobic growth of nitrate respiration, based on low gene expression levels. IMPORTANCE Understanding the assembly of iron-sulfur (Fe-S) proteins is relevant to many fields, including nitrogen fixation, photosynthesis, bioenergetics and gene regulation. Still remaining critical gaps in our knowledge are how Fe-S clusters are transferred to their target proteins and how the specificity in this process is achieved, since different forms of Fe-S clusters need to be delivered to structurally highly diverse target proteins. Numerous Fe-S carrier proteins have been identified in prokaryotes like Escherichia coli, including ErpA, IscA, SusA and NfuA. In addition, the diverse Fe-S cluster delivery proteins and their target proteins underlie a complex regulatory network of expression, to ensure that both proteins are synthesized under particular growth conditions.

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ErpA is important but not essential for the Fe/S cluster biogenesis of Escherichia coli NADH:ubiquinone oxidoreductase (complex I)

Cited by 3 publications

References 44 publications

Genetic Analysis of the ts-Lethal Mutant Δpa0665/pTS-pa0665 Reveals Its Role in Cell Morphology and Oxidative Phosphorylation in Pseudomonas aeruginosa

Genetic Analysis of the ts-Lethal Mutant Δpa0665/pTS-pa0665 Reveals Its Role in Cell Morphology and Oxidative Phosphorylation in Pseudomonas aeruginosa

The gene order in the nuo-operon is not essential for the assembly of E. coli complex I

A-Type Carrier Proteins Are Involved in [4Fe-4S] Cluster Insertion into the Radical S -Adenosylmethionine Protein MoaA for the Synthesis of Active Molybdoenzymes

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