Kinetic Buffering of Cross Talk between Bacterial Two-Component Sensors

Eli S. Groban, Elizabeth J. Clarke, Howard M. Salis, Susan M. Miller, Christopher A. Voigt

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Two-component systems are a class of sensors that enable bacteria to respond to environmental and cell-state signals. The canonical system consists of a membrane-bound sensor histidine kinase that autophosphorylates in response to a signal and transfers the phosphate to an intracellular response regulator. Bacteria typically have dozens of two-component systems. The key questions are whether these systems are linear and, if they are, how cross talk between systems is buffered. In this work, we studied the EnvZ/OmpR and CpxA/CpxR systems from Escherichia coli, which have been shown previously to exhibit slow cross talk in vitro. Using in vitro radiolabeling and a rapid quenched-flow apparatus, we experimentally measured 10 biochemical parameters capturing the cognate and non-cognate phosphotransfer reactions between the systems. These data were used to parameterize a mathematical model that was used to predict how cross talk is affected as different genes are knocked out. It was predicted that significant cross talk between EnvZ and CpxR only occurs for the triple mutant ΔompR ΔcpxA ΔactA-pta. All seven combinations of these knockouts were made to test this prediction and only the triple mutant demonstrated significant cross talk, where the cpxP promoter was induced 280-fold upon the activation of EnvZ. Furthermore, the behavior of the other knockouts agrees with the model predictions. These results support a kinetic model of buffering where both the cognate bifunctional phosphatase activity and the competition between regulator proteins for phosphate prevent cross talk in vivo.

Original languageEnglish (US)
Pages (from-to)380-393
Number of pages14
JournalJournal of Molecular Biology
Volume390
Issue number3
DOIs
StatePublished - Jul 17 2009

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Phosphates
Bacteria
Phosphoric Monoester Hydrolases
Theoretical Models
Escherichia coli
Membranes
Genes
Proteins
In Vitro Techniques
Histidine Kinase

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Cite this

Groban, Eli S. ; Clarke, Elizabeth J. ; Salis, Howard M. ; Miller, Susan M. ; Voigt, Christopher A. / Kinetic Buffering of Cross Talk between Bacterial Two-Component Sensors. In: Journal of Molecular Biology. 2009 ; Vol. 390, No. 3. pp. 380-393.
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Kinetic Buffering of Cross Talk between Bacterial Two-Component Sensors. / Groban, Eli S.; Clarke, Elizabeth J.; Salis, Howard M.; Miller, Susan M.; Voigt, Christopher A.

In: Journal of Molecular Biology, Vol. 390, No. 3, 17.07.2009, p. 380-393.

Research output: Contribution to journalArticle

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