Systematic dissection and trajectory-scanning mutagenesis of the molecular interface that ensures specificity of two-component signaling pathways

Autor: Emma A. Lubin, Barrett S. Perchuk, Jeffrey M. Skerker, Orr Ashenberg, Emily J. Capra, Michael T. Laub
Přispěvatelé: Massachusetts Institute of Technology. Computational and Systems Biology Program, Massachusetts Institute of Technology. Department of Biology, Laub, Michael T., Capra, Emily Jordan, Perchuk, Barrett, Lubin, Emma Alexandra, Ashenberg, Orr
Jazyk: angličtina
Rok vydání: 2010
Předmět:
Cancer Research
lcsh:QH426-470
Molecular Sequence Data
Regulator
Mutagenesis (molecular biology technique)
Molecular Biology/Molecular Evolution
Sequence alignment
Computational biology
Biology
Cell Biology/Cell Signaling
Substrate Specificity
Evolution
Molecular
Cell Biology/Microbial Growth and Development
Escherichia coli
Biochemistry/Cell Signaling and Trafficking Structures
Genetics
Cluster Analysis
Amino Acid Sequence
Amino Acids
Molecular Biology
Peptide sequence
Gene
Genetics (clinical)
Ecology
Evolution
Behavior and Systematics
Microbiology/Microbial Evolution and Genomics
Kinase
Escherichia coli Proteins
Computational Biology/Macromolecular Sequence Analysis
Genetics and Genomics/Microbial Evolution and Genomics
Biochemistry/Molecular Evolution
lcsh:Genetics
Evolutionary Biology/Microbial Evolution and Genomics
Mutagenesis
Phosphorylation
Signal transduction
Protein Kinases
Signal Transduction
Research Article
Zdroj: PLoS Genetics, Vol 6, Iss 11, p e1001220 (2010)
PLoS
PLoS Genetics
ISSN: 1553-7404
1553-7390
Popis: Two-component signal transduction systems enable bacteria to sense and respond to a wide range of environmental stimuli. Sensor histidine kinases transmit signals to their cognate response regulators via phosphorylation. The faithful transmission of information through two-component pathways and the avoidance of unwanted cross-talk require exquisite specificity of histidine kinase-response regulator interactions to ensure that cells mount the appropriate response to external signals. To identify putative specificity-determining residues, we have analyzed amino acid coevolution in two-component proteins and identified a set of residues that can be used to rationally rewire a model signaling pathway, EnvZ-OmpR. To explore how a relatively small set of residues can dictate partner selectivity, we combined alanine-scanning mutagenesis with an approach we call trajectory-scanning mutagenesis, in which all mutational intermediates between the specificity residues of EnvZ and another kinase, RstB, were systematically examined for phosphotransfer specificity. The same approach was used for the response regulators OmpR and RstA. Collectively, the results begin to reveal the molecular mechanism by which a small set of amino acids enables an individual kinase to discriminate amongst a large set of highly-related response regulators and vice versa. Our results also suggest that the mutational trajectories taken by two-component signaling proteins following gene or pathway duplication may be constrained and subject to differential selective pressures. Only some trajectories allow both the maintenance of phosphotransfer and the avoidance of unwanted cross-talk.
National Science Foundation (U.S.) (CAREER Award)
National Science Foundation (U.S.) (Graduate Fellowship)
Databáze: OpenAIRE
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