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Updated: Jun 8, 2026

DNA-affinity-purified Chip (DAP-chip) Method to Determine Gene Targets for Bacterial Two component Regulatory Systems
Published on: July 21, 2014
NMR assignments for the Sinorhizobium meliloti response regulator Sma0114
Sarah R Sheftic1, Preston P Garcia, Victoria L Robinson
1Department of Molecular and Cell Biology, University of Connecticut, Storrs, 06269-3125, USA.
Abstract:
Response regulators are terminal ends of bacterial two-component systems that undergo extensive structural reorganization in response to phosphoryl transfer from their cognate histidine kinases. The response regulator encoded by the gene sma0114 of Sinorhizobium meliloti is a part of a unique class of two-component systems that employ HWE histidine kinases. The distinct features of Sma0114 include a PFxFATGY motif that houses the conserved threonine in the "Y-T coupling" conformational switch which mediates output response through downstream protein-protein interactions, and the replacement of the conserved phenylalanine/tyrosine in Y-T coupling by a leucine. Here we present (1)H, (15)N, and (13)C NMR assignments for Sma0114. We identify the secondary structure of the protein based on TALOS chemical shift analysis, (3)J(HNHα) coupling constants and hydrogen-deuterium exchange. The secondary structure determined by NMR is in good agreement with that predicted from the sequence. Both methods suggest that Sma0114 differs from standard CheY-like folds by missing the fourth α-helix. Our initial NMR characterization of Sma0114 paves the way to a full investigation of the structure and dynamics of this response regulator.
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