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. 2021 Apr;27(4):513–526. doi: 10.1261/rna.077404.120

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© 2021 Wang et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article, published in RNA, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 5. — Intermediate concentrations of adapter protein are optimal for B3H signal. Effect of adapter-protein concentration on (A) MgrR–Hfq and (B) ChiX–Hfq B3H interactions. Twelve pAdapter constructs are included—six from site-directed mutagenesis (SDM) and six from forward genetic screen (mutants from screen). Data points correspond to pAdapters (left to right) 35u14, p35u9 (green), SDu7, p35u4 (blue), 35u5, pSDu5 (pink), pCW17 (white), pCW18, pCW19, pCW14–16. B3H data collected with each pAdapter are plotted against CI repression data from FW123 carrying the same construct. Both data sets were collected in the absence of IPTG. Data are fit to a quadratic function (y = ax² + bx + c; MgrR: R = 0.84; ChiX, R = 0.83). (C) Comparison of pAdapter dependence of MgrR- and ChiX–B3H interactions. Results of B3H β-gal assays performed in KB473 cells with pPrey (α-Hfq), pBaitRNA (2xMS2^hp-ChiX or 2xMS2^hp–MgrR), and pAdapter (left to right: pSDu7, p35u14, p35u5, p35u9 [green], p35u4 [blue], pSDu5 [pink]).