Fig. 3.

Activation of R. sphaeroides non-rRNA promoters in vitro with CarD_Rsp. (A) Transcription in vitro with RNAP_Rsp (20 nM) and CarD_Rsp (1,280 nM) in buffer with 100 mM NaCl (SI Appendix, Expanded Materials and Methods). Duplicate lanes with or without CarD are shown for each promoter. Transcripts derived from test promoters are indicated with red arrowheads. Gel images also show position of RNA 1 transcript from the plasmid promoter. SI Appendix, Fig. S7, shows promoter sequences. (B) Fold activation by CarD (+CarD/no CarD) for promoters shown in A whose activities were high enough to quantify accurately. SDs are shown from n = 3 to 6 assays except for ilvE1 (range from two assays). (C) Transcription in vitro of four R. sphaeroides promoters whose activities were unaffected by CarD under the same conditions as in B. SI Appendix, Fig. S7, shows promoter sequences. (D) Fold activation of promoters in C with or without CarD. The SDs shown are from n = 5 assays for ssrA, and the ranges are shown from n = 2 assays for rsp_3110 and putR. (E) WebLogo (34) representation of consensus sequence for 15 promoters whose transcription was activated by CarD in vitro (3 rRNA promoters and the 12 promoters shown in A). The −10 hexamer (−12 to −7) is boxed.