Integrative and quantitative view of the CtrA regulatory network in a stalked budding bacterium
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ABSTRACT:
PROVIDER: PRJNA554968 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR8434430.fastq.gz | Fastqsanger.gz | |||
| SRR8434431.fastq.gz | Fastqsanger.gz | |||
| SRR8434428.fastq.gz | Fastqsanger.gz | |||
| SRR8434429.fastq.gz | Fastqsanger.gz | |||
| SRR9693190.fastq.gz | Fastqsanger.gz |
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Project description:Alphaproteobacteria stand out for their complex cell cycles, which are often regulated by the DivJ/PleC-DivK-DivL-CckA-ChpT-CtrA pathway. DivJ and PleC set up the polarity of the cell, thereby eventually leading to differential activation of the DNA-binding response regulator CtrA in the two nascent daughter cells. CtrA regulates replication and transcription of many genes, thereby ensuring that processes such as motility and cell division take place at the appropriate cell cycle stage. The cell cycle of the stalked budding alphaproteobacterium Hyphomonas neptunium culminates in an asymmetric cell division at the stalk-bud junction. Here, we investigate the role of the pathway from DivJ and PleC down to CtrA in this recently established model organism. Even though DivJ and PleC are localized to opposite poles, suggesting they are involved in polarity establishment inH. neptunium, DivJ, PleC and the other components of the upstream pathway (DivK and PleD) are not essential for cell cycle regulation. In contrast, the downstream part of the pathway starting from DivL is essential and involved in the regulation of important functions such as replication inhibition, cell division and motility, as shown by the identification of the (direct) regulon of CtrA. The overlap between the regulons of DivJ and PleC, DivK and CtrA is only partial, demonstrating that additional factors feeding into the pathway must be present in H. neptunium. Furthermore, unlike in other alphaproteobacteria, the regulation of CtrA throughout the cell cycle does not take place at the level of CtrA abundance in H. neptunium. All in all, the DivL-CckA-ChpT-CtrA pathway plays an essential role in the regulation of the complicated cell cycle ofH. neptunium, but several proteins feeding into CtrA remain undiscovered. The in-depth analysis of CtrA regulation in this stalked budding organism leads to hypotheses that might also hold in well-established model organisms such as Caulobacter crescentus.
2020-03-29 | GSE124953 | GEO
Project description:The Alphaproteobacteria show a remarkable diversity of cell cycle-dependent developmental patterns, which are governed by the conserved CtrA pathway. Its central component CtrA is a DNA-binding response regulator that is controlled by a complex two-component signaling network, mediating distinct transcriptional programs in the two offspring. The CtrA pathway has been studied intensively and was shown to consist of an upstream part that reads out the developmental state of the cell and a downstream part that integrates the upstream signals and mediates CtrA phosphorylation. However, the role of this circuitry in bacterial diversification remains incompletely understood. We have therefore investigated CtrA regulation in the morphologically complex stalked budding alphaproteobacterium Hyphomonas neptunium. Compared to relatives dividing by binary fission, H. neptunium shows distinct changes in the role and regulation of various pathway components. Most notably, the response regulator DivK, which normally links the upstream and downstream parts of the CtrA pathway, is dispensable, while downstream components such as the pseudokinase DivL, the histidine kinase CckA, the phosphotransferase ChpT and CtrA are essential. Moreover, CckA is compartmentalized to the nascent bud without forming distinct polar complexes and CtrA is not regulated at the level of protein abundance. We show that the downstream pathway controls critical functions such as replication initiation, cell division and motility. Quantification of the signal flow through different nodes of the regulatory cascade revealed that the CtrA pathway is a leaky pipeline and must involve thus-far unidentified factors. Collectively, the quantitative system-level analysis of CtrA regulation in H. neptunium points to a considerable evolutionary plasticity of cell cycle regulation in alphaproteobacteria and leads to hypotheses that may also hold in well-established model organisms such as Caulobacter crescentus.
| S-EPMC7200025 | biostudies-literature
Project description:Integrative and quantitative analysis reveals proper CtrA-dependent cell cycle regulation in the absence of the DivJ/PleC asymmetry module in the stalked budding bacterium Hyphomonas neptunium
| PRJNA514449 | ENA
Project description:This SuperSeries is composed of the SubSeries listed below.
2020-03-29 | GSE134367 | GEO
Project description:Investigation of whole genome expression changes in Magnetospririllum magneticum mutants, probing the role of the CtrA regulatory pathway. The mutants are further described in a manuscript submitted for publication at J. Bacteriology. Developmental events across the prokaryotic life cycle are highly regulated at the transcriptional and post-translational levels. Key elements of a few regulatory networks are conserved among phylogenetic groups of bacteria, although the features controlled by these conserved systems are as diverse as the organisms encoding them. In this work, we probe the role of the CtrA regulatory network, conserved throughout the Alphaproteobacteria, in the magnetotactic bacterium, Magnetospirillum magneticum strain AMB-1, which possesses unique intracellular organization and compartmentalization. While we show that CtrA in AMB-1 is not essential for viability, it is required for motility, and its putative phosphorylation state dictates the ability of CtrA to activate the flagella biosynthesis gene cascade. Gene expression analysis of strains expressing active and inactive CtrA alleles point to the composition of the extended CtrA regulon, including both direct and indirect targets. These results, combined with a bioinformatic study of the AMB-1 genome, enabled the prediction of an AMB-1 specific CtrA binding site. Further, phylogenetic studies comparing CtrA sequences from Alphaproteobacteria in which the role of CtrA has been experimentally examined reveals an ancestral role of CtrA in the regulation of motility and suggests that its essential functions in other Alphaproteobacteria were acquired subsequently. Total RNA was recovered from each of the wild-type and mutant strains, reverse transcribed to cDNA, fluorescently labeled, and hybridized to whole genome microarrays. The arrays contain 7 probes/gene, with the entire genome duplicated twice. In addition, the arrays contain 7 probes for 22 unannotated ORFs and tiling of a genomic region from coordinates 977403-1097027.
2012-02-17 | E-GEOD-35625 | biostudies-arrayexpress
Project description:We wanted to test the effect on global gene expression of depleting the essential cell cycle regulator CtrA in order to determine the genes both indirectly and directly transcriptionally regulated by CtrA Gene expression changes in S. meliloti 1,2,4 and 6 hours post CtrA depletion Log-phase S. meliloti cultures carrying an IPTG-inducible allele of CtrA were split and transferred to growth media lacking IPTG (depletion) and with 1mM IPTG (control). Samples for RNA extraction were taken 1,2,4, and 6 hours after the start of the depletion experiment to monitor gene expression changes in the population after different periods of CtrA depletion.
2015-04-24 | E-GEOD-68218 | biostudies-arrayexpress
Project description:Within the bacterial phylum of the Alphaproteobacteria many species show complex life cycles. Proper regulation of these life cycles requires cell cycle regulation pathways, one of which is the so-called CtrA pathway. Key factors in the CtrA pathway are the histidine kinase CckA and the phosphotransferase ChpT, which are directly involved in cell cycle specific activation of the response regulator CtrA. Within the Alphaproteobacteria, stalked budding bacteria are even more asymmetric than most other representatives. How they regulate their cell cycle has not been studied before. Here, we investigated the transcriptional profiles of Hyphomonas neptunium cells depleted of CckA or ChpT and compared their profiles to that of wildtype cells. We identified that the essential proteins CckA and ChpT influence the transcription levels of multiple essential processes, such as cell division, as well as specific cell cycle regulated functions such as motility.
2022-01-01 | GSE134341 | GEO
Project description:We wanted to test the effect on global gene expression of depleting the essential cell cycle regulator CtrA in order to determine the genes both indirectly and directly transcriptionally regulated by CtrA Gene expression changes in S. meliloti 1,2,4 and 6 hours post CtrA depletion
2015-04-24 | GSE68218 | GEO
Project description:Within the bacterial phylum of the Alphaproteobacteria many species show complex life cycles. Proper regulation of these life cycles requires cell cycle regulation pathways, one of which is the so-called CtrA pathway. Key factors in the CtrA pathway are the histidine kinases PleC and DivJ (located at opposite poles in the cell) and the response regulator DivK. Within the Alphaproteobacteria, stalked budding bacteria are even more asymmetric than most other representatives. How they regulate their cell cycle has not been studied before. Here, we investigated the transcriptional profiles of Hyphomonas neptunium cells lacking either PleC, DivJ or DivK and compared their profiles to that of wildtype cells. We found that the changes upon deletion of DivJ, PleC and especially DivK were smaller than expected from related organisms.
2022-01-01 | GSE134343 | GEO
Project description:Investigation of whole genome expression changes in Magnetospririllum magneticum mutants, probing the role of the CtrA regulatory pathway. The mutants are further described in a manuscript submitted for publication at J. Bacteriology. Developmental events across the prokaryotic life cycle are highly regulated at the transcriptional and post-translational levels. Key elements of a few regulatory networks are conserved among phylogenetic groups of bacteria, although the features controlled by these conserved systems are as diverse as the organisms encoding them. In this work, we probe the role of the CtrA regulatory network, conserved throughout the Alphaproteobacteria, in the magnetotactic bacterium, Magnetospirillum magneticum strain AMB-1, which possesses unique intracellular organization and compartmentalization. While we show that CtrA in AMB-1 is not essential for viability, it is required for motility, and its putative phosphorylation state dictates the ability of CtrA to activate the flagella biosynthesis gene cascade. Gene expression analysis of strains expressing active and inactive CtrA alleles point to the composition of the extended CtrA regulon, including both direct and indirect targets. These results, combined with a bioinformatic study of the AMB-1 genome, enabled the prediction of an AMB-1 specific CtrA binding site. Further, phylogenetic studies comparing CtrA sequences from Alphaproteobacteria in which the role of CtrA has been experimentally examined reveals an ancestral role of CtrA in the regulation of motility and suggests that its essential functions in other Alphaproteobacteria were acquired subsequently.
2012-02-17 | GSE35625 | GEO