Genome‐wide identification of Listeria monocytogenes CodY‐binding sites
| Autor: | Boris R. Belitsky, Rajesh Kumar Biswas, Abraham L. Sonenshein |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
DNA
Bacterial Virulence Factors Operon Bacillus subtilis Microbiology Genome Article 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Coding region Binding site Molecular Biology Gene 030304 developmental biology Genetics 0303 health sciences Binding Sites biology 030306 microbiology DNase-I Footprinting Gene Expression Regulation Bacterial biology.organism_classification Listeria monocytogenes DNA-Binding Proteins chemistry DNA Protein Binding Transcription Factors |
| Zdroj: | Mol Microbiol |
| ISSN: | 1365-2958 0950-382X |
| Popis: | CodY is a global transcriptional regulator that controls, directly or indirectly, the expression of dozens of genes and operons in Listeria monocytogenes. We used in vitro DNA affinity purification combined with massively parallel sequencing (IDAP-Seq) to identify genome-wide L. monocytogenes chromosomal DNA regions that CodY binds in vitro. The total number of CodY-binding regions exceeded 2,000, but they varied significantly in their strengths of binding at different CodY concentrations. The 388 strongest CodY-binding regions were chosen for further analysis. A strand-specific analysis of the data allowed pinpointing CodY-binding sites at close to single-nucleotide resolution. Gel shift and DNase I footprinting assays confirmed the presence and locations of several CodY-binding sites. Surprisingly, most of the sites were located within genes' coding regions. The binding site within the beginning of the coding sequence of the prfA gene, which encodes the master regulator of virulence genes, has been previously implicated in regulation of prfA, but this site was weaker in vitro than hundreds of other sites. The L. monocytogenes CodY protein was functionally similar to Bacillus subtilis CodY when expressed in B. subtilis cells. Based on the sequences of the CodY-binding sites, a model of CodY interaction with DNA is proposed. |
| Databáze: | OpenAIRE |
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