Figure 5
GreA binding to the RNA polymerase. Validation of the binding activity conclusions from the fliC::lacZ activity tests (Fig. 4) by co-purification of RNAP RpoA subunit with affinity tagged GreA-His. (a) Western blots with anti-RpoA and anti-GreA-His were performed using the CF14758 strain (ΔgreA ΔgreB) containing pBb-GreA-his and pDNL278 (pGreAWT), pTrc-GreAS121P or pTrc-GreAE59G. Control Western blots showing protein abundance detected by Anti-RpoA or Anti-GreA-His in extracts before copurification are shown in Supplementary Fig. S6. Full western blots are shown in Supplementary Fig. S7. (b) RpoA abundance is normalized to His-tagged GreA abundance without IPTG. Error bars represent SD from 2 biological and technical replicates.
