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Cited by 93Bacterial Genetics and BiotechnologyIron Metabolism and DisordersEscherichia coli research studies

PfeT, a P1B4‐type ATP ase, effluxes ferrous iron and protects Bacillus subtilis against iron intoxication

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Guan, Guohua, Pinochet‐Barros, Azul, Gaballa, Ahmed, Patel, Sarju J., Argüello, José M., Helmann, John D.·2015·Molecular Microbiology
DOI 10.1111/mmi.13158PMC PMC4648274
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Abstract

Summary: Iron is an essential element for nearly all cells and limited iron availability often restricts growth. However, excess iron can also be deleterious, particularly when cells expressing high affinity iron uptake systems transition to iron rich environments. Bacillus subtilis expresses numerous iron importers, but iron efflux has not been reported. Here, we describe the B. subtilis PfeT protein (formerly YkvW/ZosA) as a P1B4‐type ATP ase in the PerR regulon that serves as an F e(II) efflux pump and protects cells against iron intoxication. Iron and manganese homeostasis in B. subtilis are closely intertwined: a pfeT mutant is iron sensitive, and this sensitivity can be suppressed by low levels of M n(II). Conversely, a pfeT mutant is more resistant to M n(II) overload. In vitro, the PfeT ATPase is activated by both F e(II) and C o(II), although only F e(II) efflux is physiologically relevant in wild‐type cells, and null mutants accumulate elevated levels of intracellular iron. Genetic studies indicate that PfeT together with the ferric uptake repressor (Fur) cooperate to prevent iron intoxication, with iron sequestration by the MrgA mini‐ferritin playing a secondary role. Protection against iron toxicity may also be a key role for related P 1B4‐type ATPases previously implicated in bacterial pathogenesis.

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  • In many bacteria, homeostasis also relies on Fe2+ export, particularly under conditions of high Fe2+ and oxidative stress.

    “In many bacteria, homeostasis also relies on Fe2+ export,136,137 particularly under conditions of high Fe2+ and oxidative stress.138−140”

    From Metals in Motion: Understanding Labile Metal Pools in Bacteria (2025) · cites this paper as [138]

    The Conclusions report that PfeT-mediated iron export helps B. subtilis survive iron intoxication and suggest that many bacteria have mechanisms to export excess iron. However, the Results and Discussion section on H2O2 stress finds that PfeT plays only a minor role in peroxide resistance, so the emphasis on Fe2+ export under oxidative stress is not supported to the same extent. (LLM, conf=0.85)

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Research works printed but not linked: 61

  1. Argüello JM Identification of ion-selectivity determinants in heavy-metal transport P1B-type ATPases. The Journal of membrane biology 2003 195 93 108 14692449
  2. Argüello JM Eren E Gónzalez-Guerrero M The structure and function of heavy metal transport P1B-ATPases. Biometals 2007 20 233 248 17219055
  3. Argüello JM Gónzalez-Guerrero M Raimunda D Bacterial transition metal P(1B)-ATPases: transport mechanism and roles in virulence. Biochemistry 2011 50 9940 9949 21999638 PMC3224801
  4. Bhubhanil S Chamsing J Sittipo P Chaoprasid P Sukchawalit R Mongkolsuk S Roles of Agrobacterium tumefaciens membrane-bound ferritin (MbfA) in iron transport and resistance to iron under acidic conditions. Microbiology 2014 160 863 871 24600024
  5. Bradford MM A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry 1976 72 248 254 942051
  6. Bsat N Herbig A Casillas-Martinez L Setlow P Helmann JD Bacillus subtilis contains multiple Fur homologues: identification of the iron uptake (Fur) and peroxide regulon (PerR) repressors. Mol Microbiol 1998 29 189 198 9701813
  7. Chen L Helmann JD Bacillus subtilis MrgA is a Dps(PexB) homologue: Evidence for Metalloregulation of an Oxidative Stress Gene. Molecular Microbiology 1995 18 295 300 8709848
  8. Chen L James LP Helmann JD Metalloregulation in Bacillus subtilis: Isolation and Characterization of Two Genes Differentially Repressed by Metal Ions. Journal of Bacteriology 1993 175 5428 5437 8396117 PMC206598
  9. Chiancone E Ceci P The multifaceted capacity of Dps proteins to combat bacterial stress conditions: Detoxification of iron and hydrogen peroxide and DNA binding. Biochimica et biophysica acta 2010 1800 798 805 20138126
  10. Cotruvo JA Jr. Stubbe J Metallation and mismetallation of iron and manganese proteins in vitro and in vivo: the class I ribonucleotide reductases as a case study. Metallomics: integrated biometal science 2012 4 1020 1036 22991063 PMC3488304
  11. Duarte V Latour JM PerR vs OhrR: selective peroxide sensing in Bacillus subtilis. Molecular bioSystems 2010 6 316 323 20094649
  12. Faulkner MJ Helmann JD Peroxide stress elicits adaptive changes in bacterial metal ion homeostasis. Antioxidants & redox signaling 2011 15 175 189 20977351 PMC3110094
  13. Faulkner MJ Ma Z Fuangthong M Helmann JD Derepression of the Bacillus subtilis PerR peroxide stress response leads to iron deficiency. J Bacteriol 2012 194 1226 1235 22194458 PMC3294777
  14. Frawley ER Crouch ML Bingham-Ramos LK Robbins HF Wang W Wright GD Fang FC Iron and citrate export by a major facilitator superfamily pump regulates metabolism and stress resistance in Salmonella Typhimurium. Proc Natl Acad Sci U S A 2013 110 12054 12059 23821749 PMC3718157
  15. Frawley ER Fang FC The ins and outs of bacterial iron metabolism. Mol Microbiol 2014 93 609 616 25040830 PMC4135372
  16. Fuangthong M Herbig AF Bsat N Helmann JD Regulation of the Bacillus subtilis fur and perR genes by PerR: not all members of the PerR regulon are peroxide inducible. J Bacteriol 2002 184 3276 3286 12029044 PMC135084
  17. Fuangthong M Helmann JD Recognition of DNA by three ferric uptake regulator (Fur) homologs in Bacillus subtilis. J Bacteriol 2003 185 6348 6357 14563870 PMC219410
  18. Gaballa A Antelmann H Aguilar C Khakh SK Song KB Smaldone GT Helmann JD The Bacillus subtilis iron-sparing response is mediated by a Fur-regulated small RNA and three small, basic proteins. Proc Natl Acad Sci U S A 2008 105 11927 11932 18697947 PMC2575260
  19. Gaballa A Helmann JD A peroxide-induced zinc uptake system plays an important role in protection against oxidative stress in Bacillus subtilis. Mol. Microbiol 2002 45 997 1005 12180919
  20. Guffanti AA Wei Y Rood SV Krulwich TA An antiport mechanism for a member of the cation diffusion facilitator family: divalent cations efflux in exchange for K+ and H+. Mol Microbiol 2002 45 145 153 12100555
  21. Gupta S Chai J Cheng J D'Mello R Chance MR Fu D Visualizing the kinetic power stroke that drives proton-coupled zinc(II) transport. Nature 2014 512 101 104 25043033 PMC4144069
  22. Harwood CR Cutting SM Molecular Biological Methods for Bacillus 1990 John Wiley and Sons, Ltd Chichester
  23. Helmann JD Specificity of Metal Sensing: Iron and Manganese Homeostasis in Bacillus subtilis. J Biol Chem 2014 289 28112 28120 25160631 PMC4192466
  24. Helmann JD Wu MF Gaballa A Kobel PA Morshedi MM Fawcett P Paddon C The global transcriptional response of Bacillus subtilis to peroxide stress is coordinated by three transcription factors. J Bacteriol 2003 185 243 253 12486061 PMC141929
  25. Herbig AF Helmann JD Roles of metal ions and hydrogen peroxide in modulating the interaction of the Bacillus subtilis PerR peroxide regulon repressor with operator DNA. Mol Microbiol 2001 41 849 859 11532148
  26. Imlay JA The molecular mechanisms and physiological consequences of oxidative stress: lessons from a model bacterium. Nature reviews. Microbiology 2013 11 443 454 23712352 PMC4018742
  27. Imlay JA The Mismetallation of Enzymes during Oxidative Stress. J Biol Chem 2014 289 28121 28128 25160623 PMC4192467
  28. Jolliffe LK Doyle RJ Streips UN The energized membrane and cellular autolysis in Bacillus subtilis. Cell 1981 25 753 763 6793239
  29. King DW Lounsbury HA Millero FJ Rates and Mechanism of Fe(II) Oxidation at Nanomolar Total Iron Concentrations. Environmental science & technology 1995 29 818 824 22200293
  30. Lanzetta PA Alvarez LJ Reinach PS Candia OA An improved assay for nanomole amounts of inorganic phosphate. Analytical biochemistry 1979 100 95 97 161695
  31. Lee JW Helmann JD Biochemical characterization of the structural Zn2+ site in the Bacillus subtilis peroxide sensor PerR. J Biol Chem 2006a 281 23567 23578 16766519
  32. Lu M Fu D Structure of the zinc transporter YiiP. Science 2007 317 1746 1748 17717154
  33. Ma Z Chandrangsu P Helmann TC Romsang A Gaballa A Helmann JD Bacillithiol is a major buffer of the labile zinc pool in Bacillus subtilis. Mol Microbiol 2014 94 756 770 25213752 PMC4227968
  34. Ma Z Faulkner MJ Helmann JD Origins of specificity and cross-talk in metal ion sensing by Bacillus subtilis Fur. Mol Microbiol 2012 86 1144 1155 23057863 PMC3508374
  35. Ma Z Lee JW Helmann JD Identification of altered function alleles that affect Bacillus subtilis PerR metal ion selectivity. Nucleic acids research 2011 39 5036 5044 21398634 PMC3130269
  36. Mandal AK Cheung WD Argüello JM Characterization of a thermophilic P-type Ag+/Cu+-ATPase from the extremophile Archaeoglobus fulgidus. J Biol Chem 2002 277 7201 7208 11756450
  37. Martin JE Waters LS Storz G Imlay JA The Escherichia coli Small Protein MntS and Exporter MntP Optimize the Intracellular Concentration of Manganese. PLoS genetics 2015 11 e1004977 25774656 PMC4361602
  38. Mascher T Hachmann AB Helmann JD Regulatory overlap and functional redundancy among Bacillus subtilis extracytoplasmic function sigma factors. J Bacteriol 2007 189 6919 6927 17675383 PMC2045236
  39. Mascher T Margulis NG Wang T Ye RW Helmann JD Cell wall stress responses in Bacillus subtilis: the regulatory network of the bacitracin stimulon. Mol Microbiol 2003 50 1591 1604 14651641
  40. McLaughlin HP Xiao Q Rea RB Pi H Casey PG Darby T Charbit A Sleator RD Joyce SA Cowart RE Hill C Klebba PE Gahan CG A putative P-type ATPase required for virulence and resistance to haem toxicity in Listeria monocytogenes. PloS one 2012 7 e30928 22363518 PMC3283593
  41. Miethke M Monteferrante CG Marahiel MA van Dijl JM The Bacillus subtilis EfeUOB transporter is essential for high-affinity acquisition of ferrous and ferric iron. Biochimica et biophysica acta 2013 1833 2267 2278 23764491
  42. Mitra B Sharma R The cysteine-rich amino-terminal domain of ZntA, a Pb(II)/Zn(II)/Cd(II)-translocating ATPase from Escherichia coli, is not essential for its function. Biochemistry 2001 40 7694 7699 11412123
  43. Moore CM Gaballa A Hui M Ye RW Helmann JD Genetic and physiological responses of Bacillus subtilis to metal ion stress. Mol Microbiol 2005 57 27 40 15948947
  44. Mostertz J Scharf C Hecker M Homuth G Transcriptome and proteome analysis of Bacillus subtilis gene expression in response to superoxide and peroxide stress. Microbiology 2004 150 497 512 14766928
  45. Neilands JB A brief history of iron metabolism. Biology of metals 1991 4 1 6 1854584
  46. Nicolaou SA Fast AG Nakamaru-Ogiso E Papoutsakis ET Overexpression of fetA (ybbL) and fetB (ybbM), Encoding an Iron Exporter, Enhances Resistance to Oxidative Stress in Escherichia coli. Applied and environmental microbiology 2013 79 7210 7219 24038693 PMC3837747
  47. Ollinger J Song KB Antelmann H Hecker M Helmann JD Role of the Fur regulon in iron transport in Bacillus subtilis. J Bacteriol 2006 188 3664 3673 16672620 PMC1482855
  48. Quadri LE Weinreb PH Lei M Nakano MM Zuber P Walsh CT Characterization of Sfp, a Bacillus subtilis phosphopantetheinyl transferase for peptidyl carrier protein domains in peptide synthetases. Biochemistry 1998 37 1585 1595 9484229
  49. Quisel JD Burkholder WF Grossman AD In vivo effects of sporulation kinases on mutant Spo0A proteins in Bacillus subtilis. J Bacteriol 2001 183 6573 6578 11673427 PMC95488
  50. Raimunda D Long JE Padilla-Benavides T Sassetti CM Argüello JM Differential roles for the Co2+ /Ni2+ transporting ATPases, CtpD and CtpJ, in Mycobacterium tuberculosis virulence. Mol Microbiol 2014 91 185 197 24255990 PMC3885230
  51. Raimunda D Long JE Sassetti CM Argüello JM Role in metal homeostasis of CtpD, a Co2+ transporting P1B4-ATPase of Mycobacterium smegmatis. Mol Microbiol 2012 84 1139 1149 22591178 PMC3370075
  52. Rensing C Fan B Sharma R Mitra B Rosen BP CopA: An Escherichia coli Cu(I)-translocating P-type ATPase. Proc Natl Acad Sci U S A 2000 97 652 656 10639134 PMC15385
  53. Rosadini CV Gawronski JD Raimunda D Argüello JM Akerley BJ A novel zinc binding system, ZevAB, is critical for survival of nontypeable Haemophilus influenzae in a murine lung infection model. Infection and immunity 2011 79 3366 3376 21576338 PMC3147578
  54. Sambrook J Fritsch EF Maniatis T Molecular Cloning: A Laboratory Manual 1990 Cold Spring Harbor Press Cold Spring Harbor
  55. Sankari S O'Brian MR A bacterial iron exporter for maintenance of iron homeostasis. J Biol Chem 2014 289 16498 16507 24782310 PMC4047416
  56. Seaver LC Imlay JA Alkyl hydroperoxide reductase is the primary scavenger of endogenous hydrogen peroxide in Escherichia coli. J Bacteriol 2001 183 7173 7181 11717276 PMC95566
  57. Smaldone GT Antelmann H Gaballa A Helmann JD The FsrA sRNA and FbpB protein mediate the iron-dependent induction of the Bacillus subtilis lutABC iron-sulfur-containing oxidases. J Bacteriol 2012a 194 2586 2593 22427629 PMC3347220
  58. Smaldone GT Revelles O Gaballa A Sauer U Antelmann H Helmann JD A global investigation of the Bacillus subtilis iron-sparing response identifies major changes in metabolism. J Bacteriol 2012b 194 2594 2605 22389480 PMC3347201
  59. Sobota JM Gu M Imlay JA Intracellular hydrogen peroxide and superoxide poison 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase, the first committed enzyme in the aromatic biosynthetic pathway of Escherichia coli. J Bacteriol 2014 196 1980 1991 24659765 PMC4010980
  60. Traore DA El Ghazouani A Jacquamet L Borel F Ferrer JL Lascoux D Ravanat JL Jaquinod M Blondin G Caux-Thang C Duarte V Latour JM Structural and functional characterization of 2-oxo-histidine in oxidized PerR protein. Nat Chem Biol 2009 5 53 59 19079268
  61. Zuber P Management of oxidative stress in Bacillus. Annual review of microbiology 2009 63 575 597 19575568

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Authors
Guan, G., Pinochet‐Barros, A., Gaballa, A., Patel, S. J., Argüello, J. M., & Helmann, J. D.
Affiliations
  • Cornell University (US)
  • Worcester Polytechnic Institute (US)
Corresponding author
Helmann, John D.
ORCID
Venue
Molecular Microbiology
Publisher
Wiley
Published
2015-09-10
Volume
98
Issue
4
Pages
787-803
ISSN
  • 0950-382X (ISSN-L)
  • 1365-2958
DOI
10.1111/mmi.13158
PMC
PMC4648274
OpenAlex
W2140545805
Cited by
93
References
7
Citations by year
2026: 5 · 2025: 6 · 2024: 10 · 2023: 7 · 2022: 11 · 2021: 8 · 2020: 13 · 2019: 5 · 2018: 3 · 2017: 14 · 2016: 10 · 2015: 1
FWCI
5.79
Type
journal article
Language
en
Open access
Licence
onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations
  • Molecular Microbiology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • NIH (GM059323)
  • China Agricultural University (100193)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Health Sciences › Medicine › Hematology › Iron Metabolism and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Escherichia coli research studies
Keywords
pFET, iron efflux, Bacillus subtilis, iron homeostasis, manganese homeostasis, fur, cobalt activation
MeSH terms
Adenosine Triphosphatases, Bacillus subtilis, Bacterial Proteins, Base Sequence, Iron, Manganese, Mutation, Repressor Proteins, Transcription Factors, Transcription, Genetic, Gene Expression Regulation, Bacterial, Regulon

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