Elucidating the CodY regulon in Staphylococcus aureus USA300 lineage using ChIPexo
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ABSTRACT:
PROVIDER: PRJNA685741 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR13259250_1.fastq.gz | Fastqsanger.gz | |||
| SRR13259250_2.fastq.gz | Fastqsanger.gz | |||
| SRR13259251_1.fastq.gz | Fastqsanger.gz | |||
| SRR13259251_2.fastq.gz | Fastqsanger.gz | |||
| SRR13259252_1.fastq.gz | Fastqsanger.gz |
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Similar Datasets
Project description:CodY is a conserved broad acting transcriptional factor that regulates the expression of genes related to amino acid metabolism and virulence in Staphylococcus aureus. CodY target genes have been studied by using in vitro DNA affinity purification and deep sequencing (IDAP-Seq). In this study we performed the first in vivo determination of CodY target genes using a novel CodY monoclonal antibody in established ChIP-exo protocols. Our results showed, 1) The same 165 CodY target genes in both strains; 2) That the differential binding intensity for the same target genes under the same conditions were due to sequence differences in the same CodY binding site in the two strains; 3) Based on transcriptomic data, a CodY regulon comprising 72 target genes that revealed that CodY is mainly involved in amino acid transport and metabolism, inorganic ion transport and metabolism, and cellular transcription and translation; and 4) CodY systematically regulated central metabolic flux to generate branched-chain amino acids (BCAAs) by mapping the CodY regulon onto a genome-scale metabolic model of S. aureus. Our study performed the first system-level analysis of CodY in two closely related dominant USA300 TCH1516 and LAC strains, thus expanding the size of the known CodY regulon, and giving new insights into the similarities and differences of CodY regulatory roles between closely related strains.
2022-08-29 | GSE163312 | GEO
Project description:CodY is a highly conserved global transcriptional factor that regulates the expression of dozens of genes related to metabolism and virulence in Staphylococcus aureus. The S. aureus CodY regulon has been studied in vitro. However, in vivo CodY DNA-binding activity and the identity of the corresponding target genes remain unknown due to lack of a ChIP-grade monoclonal antibody. Using a novel CodY monoclonal antibody that we developed in established ChIP-exo protocols, we report in vivo target genes of CodY in two S. aureus USA300 clinical strains (TCH1516 and LAC). The total number of CodY-binding sites exceeded 110, but their location varied between the two strains. The majority of the identified binding sites were located within the promoter regions. Based on the sequences of the CodY-binding sites, a model of CodY interaction with DNA is proposed. Furthermore, S. aureus CodY protein is highly conserved across a G+C Gram-positive species, including Bacillus subtilis and Listeria monocytogenes, and thus this study paves the way for exploration of the differential binding of CodY among a range of gram-positive species, including pathogenic ones
2021-08-17 | GSE159856 | GEO
Project description:Elucidation of the CodY target genes in Staphylococcus aureus USA300 using ChIPexo
| PRJNA670690 | ENA
Project description:Elucidating the BT4338 regulon
| PRJNA553806 | ENA
Project description:CodY is a conserved broad-acting transcription factor that regulates the expression of genes related to amino acid metabolism and virulence in Gram-positive bacteria. Here, we performed the first in vivo determination of CodY target genes using a novel CodY monoclonal antibody in methicillin-resistant Staphylococcus aureus (MRSA) USA300. Our results showed (i) the same 135 CodY promoter binding sites regulating the 165 target genes identified in two closely related virulent S. aureus USA300 TCH1516 and LAC strains; (ii) the differential binding intensity for the same target genes under the same conditions was due to sequence differences in the same CodY-binding site in the two strains; (iii) a CodY regulon comprising 72 target genes that are differentially regulated relative to a CodY deletion strain, representing genes that are mainly involved in amino acid transport and metabolism, inorganic ion transport and metabolism, transcription and translation, and virulence, all based on transcriptomic data; and (iv) CodY systematically regulated central metabolic flux to generate branched-chain amino acids (BCAAs) by mapping the CodY regulon onto a genome-scale metabolic model of S. aureus. Our study performed the first system-level analysis of CodY in two closely related USA300 TCH1516 and LAC strains, revealing new insights into the similarities and differences of CodY regulatory roles between the closely related strains. IMPORTANCE With the increasing availability of whole-genome sequences for many strains within the same pathogenic species, a comparative analysis of key regulators is needed to understand how the different strains uniquely coordinate metabolism and expression of virulence. To successfully infect the human host, Staphylococcus aureus USA300 relies on the transcription factor CodY to reorganize metabolism and express virulence factors. While CodY is a known key transcription factor, its target genes are not characterized on a genome-wide basis. We performed a comparative analysis to describe the transcriptional regulation of CodY between two dominant USA300 strains. This study motivates the characterization of common pathogenic strains and an evaluation of the possibility of developing specialized treatments for major strains circulating in the population.
| S-EPMC10470025 | biostudies-literature
Project description:CodY of Lactococcus lactis MG1363 is a transcriptional regulator that represses the expression of several genes encoding proteins of the proteolytic system. DNA microarray analysis, comparing the expression profiles of L. lactis MG1363 and an isogenic strain in which codY was mutated, was used to determine the CodY regulon. In peptide-rich medium and exponentially growing cells, where CodY exerts strong repressing activity, the expression of over 30 genes was significantly increased upon removal of codY. The differentially expressed genes included those predominantly involved in amino acid transport and metabolism. In addition, several genes belonging to other functional categories were derepressed, stressing the pleiotropic role of CodY. Scrutinizing the transcriptome data with bioinformatics tools revealed the presence of a novel overrepresented motif in the upstream regions of several of the genes derepressed in L. lactis MG1363codY. Evidence is presented that this 15-bps cis-sequence, AATTTTCWGAAAATT, serves as a high-affinity binding site for CodY, as shown by electrophoretic mobility shift assays and DNaseI footprinting analyses. The presence of this CodY-box is sufficient to evoke CodY-mediated regulation in vivo. A copy of this motif is also present in the upstream region of codY itself. It is shown that CodY regulates its own synthesis and requires the CodY-box and branched-chain amino acids to interact with its promoter. Keywords: genetic modification
2005-07-22 | GSE2823 | GEO
Project description:In this report, codY mutant strains were constructed and used to demonstrate the relationship of (p)ppGpp synthesized by RelP and RelQ with the activation of CodY. In addition, because CodY has not been studied in S. mutans, we used microarrays to demonstrate that this protein function as a global regulator of gene expression in S. mutans. Keywords: stress response, gene knockout analysis
2008-10-31 | GSE9641 | GEO
Project description:CodY is a widely conserved global regulator, regulating nitrogen metabolism, virulence, and stress response in Gram-positive bacteria. Here, we performed ChIP-seq to define the CodY regulon in L. lactis, and found that CodY served either as an activator or as a repressor of hundreds of genes. The genes involved in amino acid biosynthesis and transport, cell wall synthesis, nisin synthesis and immunity, and several transcription regulators were identified regulated by CodY for the first time. Intriguingly, CodY could directly bind to the downstream of codY. This study gives new insights into the function of CodY controlling cell wall synthesis, nisin synthesis and immunity, as well as self-regulation in L. lactis.
2022-01-28 | GSE139902 | GEO
Project description:The Gram-positive bacterium Staphylococcus aureus is a leading cause of severe pneumonia. Therefore, we recently investigated the fate of S. aureus upon internalization by human lung epithelial cells. This uncovered three major adaptive responses of the bacteria, involving the SigB and CodY regulons and an upregulation of the S. aureus hibernation-promoting factor (SaHPF). To explore the roles of CodY, SigB and SaHPF in the infection of human lung epithelial cells, we employed derivatives of the community-acquired MRSA strain USA300 with transposon mutations in the respective genes. Interestingly, the investigated codY mutant bacteria displayed a ‘small colony variant’-like phenotype that promotes their intracellular survival and localizes the bacteria to the host cell cytoplasm. Furthermore, our results show that CodY, SigB and saHPF contribute differentially to the different steps in the infectious process, including, host cell adhesion, invasion, intracellular survival and cytotoxicity. Notably, codY or sigB mutant bacteria experienced faster intracellular clearance compared to the parental strain, underscoring the importance of these regulators for intracellular persistence. Furthermore, we show an unprecedented role of SaHPF in human lung epithelial cell infection, including parallel effects of saHPF or SigB deficiency in adhesion and invasion. Altogether, our study focuses attention on the CodY-perceived metabolic state of the bacteria and the SigB-perceived stress in bacterial decision taking during infection. Conversely, our observations indicate important roles for the nutritional status and bacterial stress-inducing conditions in the host for the onset and subsequent course of invasive or chronic staphylococcal lung infection.
2024-09-11 | PXD044962 | Pride
Project description:CodY is a conserved regulator in gram-positive organisms described to repress metabolic genes mainly involved in nitrogen metabolism but also to control the expression of virulence genes in pathogens. We constructed codY gene-replacement mutants in three unrelated S. aureus strains (Newman, UAMS-1, RN1HG). codY mutants grew slower in a chemically defined medium compared to the wild type strains. However, only codY mutants were able to grow in medium lacking threonine. Excess of isoleucine resulted in growth inhibition in the wild type but not in the codY mutants indicating a role of isoleucine for CodY dependent repression of metabolic genes. The prototypic CodY-repressed operon ilvDBCleuABCilvA is preceded by a CodY-binding motif and repressed after up-shift with isoleucine and to a lesser extent guanidine. Transcription of the quorum sensing system agr followed a similar expression pattern. The codY dependent repression of agr is in line with the concomitant influence of CodY on typical agr regulated genes such as cap, spa fnbA and coa. However, transcriptional analysis revealed that most of these virulence genes (e.g. cap, fnbA, spa hla) are also regulated by CodY in an agr negative background. Microarray analysis revealed the large majority of codY-repressed genes are involved in amino-acid transport and metabolism, genes showing codY dependent activation were mainly involved in nucleotide transport and metabolism or virulence. In summary, CodY in S. aureus not only acts as repressor for genes involved in nitrogen metabolism but also contributes to virulence gene regulation by supporting as well as substituting agr function. The microarray was manufactured by in situ synthesis of 10'807 long oligonucleotide probes , selected as previously described (Charbonnier, 2005). It covers >98% of all ORFs annotated in strains N315 and Mu50 , MW2, COL , NCTC8325, USA300 , MRSA252 an MSSA476 including their respective plasmids.
2008-11-06 | E-GEOD-12340 | biostudies-arrayexpress