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Specific binding of the Listeria monocytogenes transcriptional regulator PrfA to target sequences requires additional factor(s) and is influenced by iron
Abstract
The PrfA protein, which is a member of the Crp/Fnr family of prokaryotic transcription activators, regulates the virulence genes of Listeria monocytogenes. In this work, specific binding of PrfA to its target DNA was determined by electrophoretic mobility‐shift assays (EMSAs) using cell‐free extracts from the two L. monocytogenes strains EGD and NCTC 7973. PrfA‐specific binding differs between the two strains, even when the concentration of PrfA was adjusted to similar levels. Both strains exhibited increased PrfA‐specific binding after a shift into minimal essential medium (MEM) without showing a significant change in the amount of PrfA protein, relative to extracts from bacteria grown in brain–heart infusion (BHI). The purified PrfA protein from strain EGD produced in Escherichia coli did not exhibit specific binding to the target DNA but did so upon addition of PrfA‐free extracts from various Listeria species and Bacillus subtilis. The observed activation of PrfA seems to be caused by a PrfA‐activating factor (Paf), which is probably a protein since elevated temperature, but not RNase treatment, destroyed the activation potential of such PrfA‐free extracts. Moreover, fractionation of these extracts by sucrose gradient centrifugation yielded the Paf activity in a fraction sedimenting at 3.2 S. Specific binding of PrfA‐containing extracts from strain EGD to the hly and actA promoter sequences was strongly inhibited by iron, whereas that of extracts from strain NCTC 7973 was only slightly reduced. The iron effect seems to be mediated by Paf rather than by PrfA itself.
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In the citing paper
“Fourteen Fur-regulated genes have been identified in L. monocytogenes, including genes that encode Fe2+ transporters and ferrichrome ABC transporters and proteins involved in Fe storage [56, 57].”
From Strategies of intracellular pathogens for obtaining iron from the environment (2015) · cites this paper as [56]
In the citing paper
“Increased Fe concentrations result in the upregulation of internalin proteins InlA and InlB, which are required for invasion [56].”
From Strategies of intracellular pathogens for obtaining iron from the environment (2015) · cites this paper as [56]
In the citing paper
“It has been shown that expression of Fur is negatively regulated by PerR, a Fur homologue that is involved in the oxidative stress response.”
From Strategies of intracellular pathogens for obtaining iron from the environment (2015) · cites this paper as [56]
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Listeria Pathogenesis and Molecular Virulence Determinants
Vázquez‐Boland, J. A., Kuhn, M., Berche, P., Chakraborty, T., Domínguez‐Bernal, G., Goebel, W., González‐Zorn, B., Wehland, J., & Kreft, J. (2001) · Clinical Microbiology Reviews · doi:10.1128/cmr.14.3.584-640.2001 · Cited by 2,147
Self-citationSchaible, U. E., & Kaufmann, S. H. E. (2004) · Nature Reviews Microbiology · doi:10.1038/nrmicro1046 · Cited by 945
“Important virulence factors that are expressed by this pathogen to facilitate bacterial intracellular invasion and spreading (including ActA, LLO and the regulator PrfA) are positively controlled by iron limitatio”
An RNA Thermosensor Controls Expression of Virulence Genes in Listeria monocytogenes
Johansson, J., Mandin, P., Renzoni, A., et al. (2002) · Cell · doi:10.1016/s0092-8674(02)00905-4 · Cited by 638
“…des Interactions Bacteries-Cellules tional activators, and activation by PrfA requires binding of PrfA to a 14 bp palindromic sequence present at PrfAInstitut Pasteur 28, rue du Docteur Roux regulated promoters, the PrfA-box (Freitag et al., 1993; Böckmann et al., 1996; Sheehan et al., 1996).”
Dussurget, O., Cabanes, D., Dehoux, P., Lecuit, M., Buchrieser, C., Glaser, P., & Cossart, P. (2002) · Molecular Microbiology · doi:10.1046/j.1365-2958.2002.03080.x · Cited by 356
“, 1997) and iron concentration (Bockmann et al., 1996).”
BackgroundAlmeida, R. S., Brunke, S., Albrecht, A., Thewes, S., Laue, M., Edwards, J. E., Filler, S. G., & Hube, B. (2008) · PLoS Pathogens · doi:10.1371/journal.ppat.1000217 · Cited by 330
“For example, the expression of virulence genes in Listeria monocytogenes was found to be positively controlled by iron limitation [23] and infections with Mycobacterium tuberculosis were reported to be more fatal when iron was accessible [24].”
Background
Metadata
- Authors
- Böckmann, R., Dickneite, C., Middendorf, B., Goebel, W., & Sokolovic, Z.
- Affiliations
- University of Würzburg (DE)
- Corresponding author
- Goebel, Werner
- ORCID
- Goebel, Werner
- Venue
- Molecular Microbiology
- Publisher
- Wiley
- Published
- 1996-11
- Volume
- 22
- Issue
- 4
- Pages
- 643-653
- ISSN
- 0950-382X (ISSN-L)
- 1365-2958
- OpenAlex
- W2122365028
- Cited by
- 101
- References
- 0
- Citations by year
- 2026: 1 · 2024: 1 · 2022: 3 · 2021: 2 · 2020: 2 · 2019: 4 · 2018: 1 · 2017: 2 · 2016: 2 · 2015: 5 · 2014: 2 · 2012: 3
- FWCI
- 5.68
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Molecular Microbiology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Universität Ulm
- University of California, Davis
- Topics
- Life Sciences › Agricultural and Biological Sciences › Food Science › Listeria monocytogenes in Food Safety
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
- Keywords
- PrfA, Listeria monocytogenes, transcriptional regulation, DNA binding, transcriptional activator, iron regulation, virulence gene regulation, strain differences, protein-DNA interactions
- MeSH terms
- Bacterial Proteins, Bacterial Toxins, Culture Media, DNA, Bacterial, Escherichia coli, Heat-Shock Proteins, Hemolysin Proteins, Humans, Iron, Listeria monocytogenes, Membrane Proteins, Peptide Termination Factors, Promoter Regions, Genetic, Recombinant Fusion Proteins, Trans-Activators, Gene Expression
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