RNAP can restart transcription elongation after transcript cleavage by MtbGre.
The starting materials in all the experiments were purified stalled complexes with radiolabeled RNA. (A) Cleavage-restart assay on T7A1-TEC. The scheme on the left depicts the reaction process. The TEC was incubated without (lane 1) or with MtbGre (lane 2 and 3), or NTPs (lane 3 and 4), as indicated. (B) Similar assay was carried out on M. smegmatis PrnB promoter template (the RNAP stalls at +39 position in the absence of UTP in the reaction mix). Transcripts were analyzed by resolving on 20% urea PAGE.