Requirement of both kinase and phosphatase activities of an Escherichia coli receptor (Taz1) for ligand-dependent signal transduction

Yang Yang, Masayori Inouye

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63 Citations (Scopus)

Abstract

Taz1 is a hybrid receptor in the Escherichia coli cytoplasmic membrane, consisting of the N-terminal ligand binding domain of Tar (a chemoreceptor for aspartate) and the C-terminal signaling domain EnvZ (an osmosensor). The binding of aspartate to an extra cytoplasmic domain induces the transmembrane signal to the cytoplasmic signaling domain. The signaling domain functioning as a protein kinase evokes a response by transferring a phosphate from an intracellular histidine to OmpR. This domain also encodes an OmpR-specific phosphatase whose action is crucial in completing the OmpR phosphorylation cycle. Phosphorylated OmpR acts as a transcriptional activator for the ompC gene. A number of mutations were introduced into the signaling domain in conserved sequences of the prokaryotic histidine kinase family. All Taz1 mutants lost the ability to both autophosphorylate the histidine residue and transfer the phosphate to OmpR. These mutated receptors were unable to activate ompC-lacZ expression. However, ompC-lacZ was able to be activated by complementation of Taz1 mutants. In some combinations, two different defective Taz1 mutants could restore both OmpR kinase and phosphatase activities when co-expressed. In other combinations only kinase activity was restored. Aspartate-inducible ompC-lacZ expression was restored only in the former cases, while in the latter cases ompC-lacZ expression became constitutive. These results indicate that the kinase activity is essential to activate ompC expression while the phosphatase activity is required to regulate ompC gene expression in a ligand-dependent manner.

Original languageEnglish (US)
Pages (from-to)335-342
Number of pages8
JournalJournal of molecular biology
Volume231
Issue number2
DOIs
StatePublished - May 20 1993

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Phosphoric Monoester Hydrolases
Aspartic Acid
Signal Transduction
Phosphotransferases
Escherichia coli
Ligands
Histidine
Phosphates
Tars
Conserved Sequence
Protein Kinases
Phosphorylation
Cell Membrane
Gene Expression
Mutation
Genes

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Keywords

  • EnvZ
  • Kinase
  • Phosphatase
  • Signal transduction
  • Tar

Cite this

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abstract = "Taz1 is a hybrid receptor in the Escherichia coli cytoplasmic membrane, consisting of the N-terminal ligand binding domain of Tar (a chemoreceptor for aspartate) and the C-terminal signaling domain EnvZ (an osmosensor). The binding of aspartate to an extra cytoplasmic domain induces the transmembrane signal to the cytoplasmic signaling domain. The signaling domain functioning as a protein kinase evokes a response by transferring a phosphate from an intracellular histidine to OmpR. This domain also encodes an OmpR-specific phosphatase whose action is crucial in completing the OmpR phosphorylation cycle. Phosphorylated OmpR acts as a transcriptional activator for the ompC gene. A number of mutations were introduced into the signaling domain in conserved sequences of the prokaryotic histidine kinase family. All Taz1 mutants lost the ability to both autophosphorylate the histidine residue and transfer the phosphate to OmpR. These mutated receptors were unable to activate ompC-lacZ expression. However, ompC-lacZ was able to be activated by complementation of Taz1 mutants. In some combinations, two different defective Taz1 mutants could restore both OmpR kinase and phosphatase activities when co-expressed. In other combinations only kinase activity was restored. Aspartate-inducible ompC-lacZ expression was restored only in the former cases, while in the latter cases ompC-lacZ expression became constitutive. These results indicate that the kinase activity is essential to activate ompC expression while the phosphatase activity is required to regulate ompC gene expression in a ligand-dependent manner.",
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T1 - Requirement of both kinase and phosphatase activities of an Escherichia coli receptor (Taz1) for ligand-dependent signal transduction

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AU - Inouye, Masayori

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N2 - Taz1 is a hybrid receptor in the Escherichia coli cytoplasmic membrane, consisting of the N-terminal ligand binding domain of Tar (a chemoreceptor for aspartate) and the C-terminal signaling domain EnvZ (an osmosensor). The binding of aspartate to an extra cytoplasmic domain induces the transmembrane signal to the cytoplasmic signaling domain. The signaling domain functioning as a protein kinase evokes a response by transferring a phosphate from an intracellular histidine to OmpR. This domain also encodes an OmpR-specific phosphatase whose action is crucial in completing the OmpR phosphorylation cycle. Phosphorylated OmpR acts as a transcriptional activator for the ompC gene. A number of mutations were introduced into the signaling domain in conserved sequences of the prokaryotic histidine kinase family. All Taz1 mutants lost the ability to both autophosphorylate the histidine residue and transfer the phosphate to OmpR. These mutated receptors were unable to activate ompC-lacZ expression. However, ompC-lacZ was able to be activated by complementation of Taz1 mutants. In some combinations, two different defective Taz1 mutants could restore both OmpR kinase and phosphatase activities when co-expressed. In other combinations only kinase activity was restored. Aspartate-inducible ompC-lacZ expression was restored only in the former cases, while in the latter cases ompC-lacZ expression became constitutive. These results indicate that the kinase activity is essential to activate ompC expression while the phosphatase activity is required to regulate ompC gene expression in a ligand-dependent manner.

AB - Taz1 is a hybrid receptor in the Escherichia coli cytoplasmic membrane, consisting of the N-terminal ligand binding domain of Tar (a chemoreceptor for aspartate) and the C-terminal signaling domain EnvZ (an osmosensor). The binding of aspartate to an extra cytoplasmic domain induces the transmembrane signal to the cytoplasmic signaling domain. The signaling domain functioning as a protein kinase evokes a response by transferring a phosphate from an intracellular histidine to OmpR. This domain also encodes an OmpR-specific phosphatase whose action is crucial in completing the OmpR phosphorylation cycle. Phosphorylated OmpR acts as a transcriptional activator for the ompC gene. A number of mutations were introduced into the signaling domain in conserved sequences of the prokaryotic histidine kinase family. All Taz1 mutants lost the ability to both autophosphorylate the histidine residue and transfer the phosphate to OmpR. These mutated receptors were unable to activate ompC-lacZ expression. However, ompC-lacZ was able to be activated by complementation of Taz1 mutants. In some combinations, two different defective Taz1 mutants could restore both OmpR kinase and phosphatase activities when co-expressed. In other combinations only kinase activity was restored. Aspartate-inducible ompC-lacZ expression was restored only in the former cases, while in the latter cases ompC-lacZ expression became constitutive. These results indicate that the kinase activity is essential to activate ompC expression while the phosphatase activity is required to regulate ompC gene expression in a ligand-dependent manner.

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