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Cited by 254Bacterial Genetics and BiotechnologyBacterial biofilms and quorum sensingIron Metabolism and Disorders

Gene repression by the ferric uptake regulator in Pseudomonas aeruginosa: cycle selection of iron-regulated genes.

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Ochsner, U A, Vasil, M L·1996·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.93.9.4409PMC PMC39551
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Abstract

The expression of at least 24 distinct genes of Pseudomonas aeruginosa PAO1 is under direct control of the "ferric uptake regulator" (Fur). Novel targets of the Fur protein were isolated in a powerful SELEX (systematic evolution of ligands by exponential enrichment)-like cycle selection consisting of in vitro DNA-Fur interaction, binding to anti-Fur antibody, purification on protein G, and PCR amplification. DNA fragments obtained after at least three exponential enrichment cycles were cloned and subjected to DNA mobility-shift assays and DNase I footprint analyses to verify the specific interaction with the Fur protein in vitro. Iron-dependent expression of the corresponding genes in vivo was monitored by RNase protection analysis. In total, 20 different DNA fragments were identified which represent actual Pseudomonas iron-regulated genes (PIGs). While four PIGs are identical to already known genes (pfeR, pvdS, tonB, and fumC, respectively), 16 PIGs represent previously unknown genes. Homology studies of the putative proteins encoded by the PIGs allowed us to speculate about their possible function. Two PIG products were highly similar to siderophore receptors from various species, and three PIG products were significantly homologous to alternative sigma factors. Furthermore, homologs of the Escherichia coli ORF1-tolQ, nuoA, stringent starvation protein Ssp, and of a two-component regulatory system similar to the Pseudomonas syringae LemA sensor kinase were identified. The putative gene products of seven additional PIGs did not show significant homologies to any known proteins. The PIGs were mapped on the P.aeruginosa chromosome. Their possible role in iron metabolism and virulence of P. aeruginosa is discussed.

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Authors
Ochsner, U. A., & Vasil, M. L.
Affiliations
  • University of Colorado Health (US)
  • Harvard University (US)
ORCID
Ochsner, U A
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
1996-04-30
Volume
93
Issue
9
Pages
4409-4414
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.93.9.4409
PMC
PMC39551
OpenAlex
W2124372396
Cited by
254
References
17
Citations by year
2026: 3 · 2025: 2 · 2024: 3 · 2023: 6 · 2022: 12 · 2021: 4 · 2020: 6 · 2019: 5 · 2018: 8 · 2017: 4 · 2016: 7 · 2015: 6 · 2014: 10 · 2013: 9 · 2012: 7
FWCI
4.40
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Science Foundation
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
  • National Institutes of Health
  • National Institute of Allergy and Infectious Diseases (AI15940)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
  • Health Sciences › Medicine › Hematology › Iron Metabolism and Disorders
Keywords
ferric uptake regulator, Fur protein, Pseudomonas aeruginosa PAO1, gene repression, protein-DNA interactions, siderophore receptors, alternative sigma factor, two-component systems, iron metabolism, virulence
MeSH terms
Amino Acid Sequence, Bacterial Proteins, Base Sequence, DNA, Bacterial, Molecular Sequence Data, Pseudomonas aeruginosa, Repressor Proteins, Substrate Specificity, Restriction Mapping, Gene Expression Regulation, Bacterial, Polymerase Chain Reaction, Open Reading Frames, Sequence Homology, Amino Acid, DNA Primers, Helix-Turn-Helix Motifs

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