title: Sensor-response regulator interactions in a cross-regulated signal transduction network.
abstract: Two-component signal transduction involves phosphoryl transfer between a histidine kinase sensor and a response regulator effector. The nitrate-responsive two-component signal transduction systems in Escherichia coli represent a paradigm for a cross-regulation network, in which the paralogous sensor-response regulator pairs, NarX-NarL and NarQ-NarP, exhibit both cognate (e.g. NarX-NarL) and non-cognate (e.g. NarQ-NarL) interactions to control output. Here, we describe results from bacterial adenylate cyclase two-hybrid (BACTH) analysis to examine sensor dimerization as well as interaction between sensor-response regulator cognate and non-cognate pairs. Although results from BACTH analysis indicated that the NarX and NarQ sensors interact with each other, results from intragenic complementation tests demonstrate that they do not form functional heterodimers. Additionally, intragenic complementation shows that both NarX and NarQ undergo intermolecular autophosphorylation, deviating from the previously reported correlation between DHp (dimerization and histidyl phosphotransfer) domain loop handedness and autophosphorylation mode. Results from BACTH analysis revealed robust interactions for the NarX-NarL, NarQ-NarL and NarQ-NarP pairs but a much weaker interaction for the NarX-NarP pair. This demonstrates that asymmetrical cross-regulation results from differential binding affinities between different sensor-regulator pairs. Finally, results indicate that the NarL effector (DNA-binding) domain inhibits NarX-NarL interaction. Missense substitutions at receiver domain residue Ser-80 enhanced NarX-NarL interaction, apparently by destabilizing the NarL receiver-effector domain interface.
authors: 1​ Food Science Graduate Group, University of California, Davis, CA, 95616-8665, USA.
authors: TuAnh Ngoc
authors: TN
authors: Huynh
authors: 2​ Department of Microbiology & Molecular Genetics, University of California, Davis, CA 95616-8665, USA.
authors: Li-Ling
authors: LL
authors: Chen
authors: 2​ Department of Microbiology & Molecular Genetics, University of California, Davis, CA 95616-8665, USA 1​ Food Science Graduate Group, University of California, Davis, CA, 95616-8665, USA.
authors: Valley
authors: V
authors: Stewart
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D053263:Gene Regulatory Networks; D005816:Genetic Complementation Test; D010766:Phosphorylation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
chemical_list: D029968:Escherichia coli Proteins