Site-directed mutagenesis identifies a molecular switch involved in copper sensing by the histidine kinase CinS in Pseudomonas putida KT2440.
In the presence of copper, Pseudomonas putida activates transcription of cinAQ via the two-component system CinS-CinR. The CinS-CinR TCS was responsive to 0.5 microM copper and was specifically activated only by copper and silver. Modeling studies of CinS identified a potential copper binding site containing H37 and H147. CinS mutants with H37R and H147R mutations had an almost 10-fold reduced copper-dependent induction of cinAQ compared to the wild type.
Quaranta
Davide
D
Department of Soil, Water, and Environmental Science, University of Arizona, Tucson, AZ 85721, USA.
McEvoy
Megan M
MM
Rensing
Christopher
C
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D001665:Binding Sites / Q000235:genetics / Q000502:physiology; D003300:Copper / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000235:genetics; D000071677:Histidine Kinase; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D016958:Pseudomonas putida / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D014131:Trace Elements / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics
D001426:Bacterial Proteins; D014131:Trace Elements; D003300:Copper; D011494:Protein Kinases; D000071677:Histidine Kinase