ArticleProceedings of the National Academy of Sciences of the United States of America2013
Probing kinase and phosphatase activities of two-component systems in vivo with concentration-dependent phosphorylation profiling.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 49 papers.
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Who cites it
49 citing papers in PubMed, 78 citations in OpenAlex.
- Multifaceted Evolution of the PhoPQ Two-Component System in Salmonella enterica Enhanced the Expression of Horizontally Acquired Virulence Genes.Molecular microbiology · 2025Article
- Constitutive activation of two-component systems reveals regulatory network interactions in Streptococcus agalactiae.Nature communications · 2024Article
- Experimental measurement and computational prediction of bacterial Hanks-type Ser/Thr signaling system regulatory targets.Molecular microbiology · 2024Review
- Chemical Reaction Models in Synthetic Promoter Design in Bacteria.Methods in molecular biology (Clifton, N.J.) · 2024Article
- CpxA/R-Controlled Nitroreductase Expression as Target for Combinatorial Therapy against Uropathogens by Promoting Reactive Oxygen Species Generation.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- Sequestration of histidine kinases by non-cognate response regulators establishes a threshold level of stimulation for bacterial two-component signaling.Nature communications · 2023Article
- Natural flavonoids disrupt bacterial iron homeostasis to potentiate colistin efficacy.Science advances · 2023Article
- Genome-wide phage susceptibility analysis in Acinetobacter baumannii reveals capsule modulation strategies that determine phage infectivity.PLoS pathogens · 2023Article
- Exploring the mono-/bistability range of positively autoregulated signaling systems in the presence of competing transcription factor binding sites.PLoS computational biology · 2022Article
- Real-time detection of response regulator phosphorylation dynamics in live bacteria.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Acinetobacter baumannii Regulates Its Stress Responses via the BfmRS Two-Component Regulatory System.Journal of bacteriology · 2022Article
- A balancing act in transcription regulation by response regulators: titration of transcription factor activity by decoy DNA binding sites.Nucleic acids research · 2021Article
- Structural insight into the role of the PAS domainfor signal transduction in sensor-kinase BvgS.Journal of bacteriology · 2021Article
- From sequence to information.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2020Review
- Evidence of Robustness in a Two-Component System Using a Synthetic Circuit.Journal of bacteriology · 2020Article
- The Staphylococcus aureus ArlRS two-component system regulates virulence factor expression through MgrA.Molecular microbiology · 2020Article
- Control of theEcoSal Plus · 2019Review
- Quantifying dynamic mechanisms of auto-regulation in Escherichia coli with synthetic promoter in response to varying external phosphate levels.Scientific reports · 2019Article
- Overcoming the Cost of Positive Autoregulation by Accelerating the Response with a Coupled Negative Feedback.Cell reports · 2018Article
- Phosphatase activity tunes two-component system sensor detection threshold.Nature communications · 2018Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
Quantitative analyses of protein concentrations, modifications and activities in their native environments are playing an increasingly vital role in unraveling the general principles underlying signal transduction pathways. The prevalent bacterial two-component systems (TCSs) use a central phosphotransfer for signaling; however, in vivo characterization of the kinase and phosphatase activities of TCS proteins is often limited by traditional transcriptional reporter assays and complicated by simultaneous actions of multiple TCS activities. Here, we report a strategy that combines concentration-dependent phosphorylation profiling and mathematical modeling to characterize the cellular activities of the archetype Escherichia coli PhoR/PhoB system. Phosphorylation of the response regulator (RR) PhoB has been found to be dependent on the total concentrations of PhoB/PhoR and saturated at high concentrations. The relationship between RR phosphorylation and total concentrations has been defined by the modeling of the kinase and phosphatase reactions and quantified to derive the biochemical parameters of the PhoR/PhoB system in vivo. In a further test of this approach on a PhoB mutant, PhoB(F20D), it proved highly effective in exploring the mechanistic differences of TCSs that are not revealed by traditional reporter assays. Measurement of biochemical parameters for PhoB(F20D) led to the discovery that a weaker interaction between the histidine sensor kinase and RR could result in a higher and nonrobust phosphorylation due to diminished phosphatase activities.
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