期刊
PROTEIN SCIENCE
卷 11, 期 11, 页码 2644-2654出版社
COLD SPRING HARBOR LAB PRESS
DOI: 10.1110/ps.0220402
关键词
CheY; Escherichia coli; random mutagenesis; two-component systems; chemotaxis
资金
- NIGMS NIH HHS [R01 GM57773, F32 GM64228, R01 GM057773, F32 GM064228] Funding Source: Medline
Response regulator proteins of two-component systems are usually activated by phosphorylation. The phosphorylated response regulator protein CheYsimilar toP mediates the chemotaxis response in Escherichia coli. We performed random mutagenesis and selected CheY mutants that are constitutively active in the absence of phosphorylation. Although a single amino acid substitution can lead to constitutive activation, no single DNA base change can effect such a transition. Numerous different sets of mutations that activate in synergy were selected in several different combinations. These mutations were all located on the side of CheY defined by alpha4, beta5, alpha5, and alpha1. Our findings argue against the two-state hypothesis for response regulator activation. We propose an alternative intermolecular mechanism that involves a dynamic interplay between response regulators and their effector targets.
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