ClbR Is the Key Transcriptional Activator of Colibactin Gene Expression in Escherichia coli

Autor: Jörg Overmann, Boyke Bunk, Martina Selle, Daniel Sauer, Nadège Bossuet-Greif, Ulrich Dobrindt, Simone König, Marina Brinkmann, Alexander Wallenstein, Cathrin Spröer, Stefan Homburg, Haleluya Wami, Jean-Philippe Nougayrède, Rudolf von Bünau, Eric Oswald, Rolf Müller, Nadine Rehm
Přispěvatelé: Institute of Hygiene, University of Münster, Università cattolica del Sacro Cuore [Roma] (Unicatt), Interdisciplinary Center for Clinical Research, Medical Faculty of Münster, Institute of Molecular Infection Biology, University of Würzburg, Institut de Recherche en Santé Digestive (IRSD ), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Helmholtz Institute for Pharmaceutical Research Saarland, Helmholtz Centre for Infection Research and Department of Pharmaceutical Biotechnology, Saarland University, German Collection of Microorganismes and Cell Cultures (DSMZ), DSMZ, German Center for Infection Research - partner site Hannover-Braunschweig (DZIF), Pharma Zentrale GmbH, Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zelllkulturen GmBH - DSMZ (GERMANY), the Interdisciplinary Center for Clinical Research of the Medical Faculty Münster (Dob2/013/12), the German Research Foundation (DO789/11-1), the German Research Foundation (SFB 479, TP A1), SEGUIN, Nathalie, Università cattolica del Sacro Cuore = Catholic University of the Sacred Heart [Roma] (Unicatt), Westfälische Wilhelms-Universität Münster = University of Münster (WWU), HIPS, Helmholtz-Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1 66123 Saarbrücken, Germany.
Jazyk: angličtina
Rok vydání: 2020
Předmět:
VNTR
lcsh:QR1-502
secondary metabolite
Biology
010402 general chemistry
[SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry
Molecular Biology/Molecular biology
01 natural sciences
Microbiology
lcsh:Microbiology
cytopathic effect
Transcriptome
03 medical and health sciences
Polyketide
polyketide
Transcription (biology)
Nonribosomal peptide
[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Genomics [q-bio.GN]
Gene expression
Molecular Biology
030304 developmental biology
Regulation of gene expression
chemistry.chemical_classification
0303 health sciences
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Molecular biology
Pathogenicity island
QR1-502
0104 chemical sciences
3. Good health
Cell biology
chemistry
Proteome
[SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry
Molecular Biology/Genomics [q-bio.GN]
RNA-seq
Zdroj: MSphere
MSphere, American Society for Microbiology., 2020, 5 (4), ⟨10.1128/mSphere.00591-20⟩
MSphere, 2020, 5 (4), ⟨10.1128/mSphere.00591-20⟩
mSphere, Vol 5, Iss 4 (2020)
mSphere
United States
mSphere, Vol 5, Iss 4, p e00591-20 (2020)
ISSN: 2379-5042
DOI: 10.1128/mSphere.00591-20⟩
Popis: Colibactin is a nonribosomal peptide/polyketide hybrid natural product expressed by different members of the Enterobacteriaceae which can be correlated with induction of DNA double-strand breaks and interference with cell cycle progression in eukaryotes. Regulatory features of colibactin expression are only incompletely understood. We used Escherichia coli strain M1/5 as a model to investigate regulation of expression of the colibactin determinant at the transcriptional level and to characterize regulatory elements located within the colibactin pathogenicity island itself. We measured clbR transcription in vitro and observed that cultivation in defined minimal media led to increased colibactin expression relative to rich media. Transcription of clbR directly responds to iron availability. We also characterized structural DNA elements inside the colibactin determinant involved in ClbR-dependent regulation, i.e., ClbR binding sites and a variable number of tandem repeats located upstream of clbR. We investigated the impact of clbR overexpression or deletion at the transcriptome and proteome levels. Moreover, we compared global gene regulation under these conditions with that occurring upon overexpression or deletion of clbQ, which affects the flux of colibactin production. Combining the results of the transcriptome and proteome analyses with indirect measurements of colibactin levels by cell culture assays and an approximate quantification of colibactin via the second product of colibactin cleavage from precolibactin, N-myristoyl-d-asparagine, we demonstrate that the variable number of tandem repeats plays a significant regulatory role in colibactin expression. We identify ClbR as the only transcriptional activator known so far that is specific and essential for efficient regulation of colibactin production. IMPORTANCE The nonribosomal peptide/polyketide hybrid colibactin can be considered a bacterial virulence factor involved in extraintestinal infection and also a procarcinogen. Nevertheless, and despite its genotoxic effect, colibactin expression can also inhibit bacterial or tumor growth and correlates with probiotic anti-inflammatory and analgesic properties. Although the biological function of this natural compound has been studied extensively, our understanding of the regulation of colibactin expression is still far from complete. We investigated in detail the role of regulatory elements involved in colibactin expression and in the growth conditions that promote colibactin expression. In this way, our data shed light on the regulatory mechanisms involved in colibactin expression and may support the expression and purification of this interesting nonribosomal peptide/polyketide hybrid for further molecular characterization.
Databáze: OpenAIRE
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