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. Author manuscript; available in PMC: 2019 Jan 1.
Published in final edited form as: Mol Microbiol. 2017 Dec 8;107(2):164–179. doi: 10.1111/mmi.13868

Figure 3. Hydrogen bonding in the α11-β5 loop stabilizes the inactive state of PhyR.

Figure 3

(A) Phosphorylation–induced rearrangement of α11-β5 loop hydrogen bonds of PhyR (top) and PhyR~P following loop rearrangement (bottom) observed in select MD simulations. Sidechain–sidechain and sidechain–backbone hydrogen bonds are colored red and blue, respectively. (B) Bacteria two-hybrid (BTH) interaction assays with PhyR α11-β5 loop alanine mutants. Inset: Cartoon outlining BTH interaction assay. (C) BTH interaction assays for L227A, T228A or E233A combined with the D192A (non-phosphorylatable) PhyR mutation. (D) Activity of a GSR transcriptional reporter (PsigU) in the absence (blue) or presence (red) of hyperosmotic stress. Inset: Cartoon outlining transcriptional reporter scheme. Controls for BTH and PsigU assays are summarized in Figure S7. Error bars represent standard deviation.