Skip to main content
. Author manuscript; available in PMC: 2023 Jul 11.
Published in final edited form as: Nature. 2023 Jan 11;613(7945):783–789. doi: 10.1038/s41586-022-05604-1

Extended Data Fig. 3. Disruption of NT −4 pocket interaction does not affect termination efficiency or pre-termination pausing.

Extended Data Fig. 3

a. NT −4 pocket with R180, R465 and V469 shown as sticks. An adenine on the non-template DNA is flipped into the pocket and hydrogen bonds with R465. b. DNA template encoding λPR promoter and λtR2 terminator used for promoter-initiated, in vitro transcription assay of the effects of pocket mutants on termination. c. Effect of pocket mutants on the termination efficiency of wild-type λtR2 terminator. d. Effect of pocket mutants on the termination efficiency of a λtR2 terminator containing a different downstream DNA. e. Effect of pocket mutants on the termination efficiency of ϕt500 terminator. f. Effect of pocket mutants on the termination efficiency of a scaffold-based λtR2 terminator in the absence or presence of E. coli NusA. Data are presented as mean ± SD, n=3 biologically independent experiments for c-f. g. Effect of pocket mutants on pre-termination pausing at U7 and U8. A variant of λtR2 terminator lacking the upstream half of the terminator hairpin (pRM1234; Supplementary Table 1) and an antisense DNA to prevent backtracking of G33 complexes were used. See legend to Extended Data Fig. 8c and methods for more details. Results from a single replicate are shown. Raw data for the gels in this figure can be found in Supplementary Fig. 1a-d.