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. 2001 Jan 15;29(2):423–429. doi: 10.1093/nar/29.2.423

Figure 2.

Figure 2

Termination facilitates reinitiation. (A) Time course of transcription. Assays contained 10 ng template DNA (pMrT2/EcoRI) and partially purified transcription factors. Reactions were started by addition of nucleotides and samples were taken at the time points indicated. (B) Sarkosyl sensitivity of TTF-I-mediated transcription stimulation. Transcription reactions contained 6 µl S-100 extracts and 30 ng pMrT2/EcoRI in the absence (lanes 1 and 3) or presence of 5 fmol recombinant TTFΔN185 (lanes 2 and 4). To prevent reinitiation, the reactions shown in lanes 3 and 4 contained 0.045% Sarkosyl. (C) TTF-I is not sufficient for the enhanced synthesis of terminated transcripts. Transcriptions were performed in a reconstituted system containing 20 ng pMrT2/EcoRI, 3 ng recombinant UBF, purified TIF-IA, TIF-IB, TIF-IC and two preparations of Pol I, i.e., H-400 (lanes 1 and 2) or S-300 (lanes 3 and 4). Where indicated, 5 fmol TTFΔN185 were added.