title: The DNA-binding domain of the Escherichia coli CpxR two-component response regulator is constitutively active and cannot be fully attenuated by fused adjacent heterologous regulatory domains.
abstract: Two-component systems (TCS) based on a sensor histidine kinase and a phosphorylated cognate target regulator allow rapid responses to environmental changes. TCS are highly evolutionarily conserved, though in only a few cases are the inducing signals understood. This study focuses on the Escherichia coli CpxR response regulator that responds to periplasmic and outer-membrane stress. N-terminal deletion mutations have been isolated that render the transcription factor constitutively active, indicating that the N terminus functions, in part, to keep the C-terminal winged-helix DNA-binding effector domain in an inactive state. Analysis of truncations spanning the CpxR interdomain region revealed that mutants retaining the alpha5 helix significantly augment activation. Hybrid proteins obtained by fusing the CpxR effector domain to structurally similar heterologous N-terminal regulatory domains, or even GFP, failed to restore repression to the C-terminal domain. These findings shed light on the mechanism of CpxR effector domain activation and on the investigation of constitutive mutants obtained by truncation in other TCS.
authors.affiliation: Division of Infectious Diseases, Geneva University Hospital, 24 Micheli-du-Crest, CH-1211 Geneva 14, Switzerland.
authors.forename: Caroline
authors.initials: C
authors.lastname: Tapparel
authors.affiliation: Division of Infectious Diseases, Geneva University Hospital, 24 Micheli-du-Crest, CH-1211 Geneva 14, Switzerland.
authors.forename: Antoinette
authors.initials: A
authors.lastname: Monod
authors.affiliation: Division of Infectious Diseases, Geneva University Hospital, 24 Micheli-du-Crest, CH-1211 Geneva 14, Switzerland.
authors.forename: William L
authors.initials: WL
authors.lastname: Kelley
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000502:physiology*; D004926:Escherichia coli / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D005809:Genes, Regulator; D009154:Mutation; D011494:Protein Kinases / Q000737:chemistry / Q000502:physiology*; D017434:Protein Structure, Tertiary
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C085504:CpxR protein, Bacteria; D011494:Protein Kinases