Fig. 6
TTD mutations alter CAK release and Pol II phosphorylation. a Biotinylated AdMLP bound to streptavidin magnetic beads was incubated with Pol II, TFIIA, TFIIB, TFIID (TBP), TFIIF, and TFIIH/WT, in presence (when indicated, +) of ATP (200 μM) and either rIIEαβ/WT, /A150P, or /D187Y. The immunoblot signals for TFIIEα, CDK7, and CyclinH were quantified and plotted in arbitrary units (a.u.). The results (n = 3, means ± s.d.) are representative of three independent experiments. b Streptavidin magnetic beads were incubated (when indicated, +) with biotinylated AdMLP, Pol II, TFIIA, TFIIB, TFIID, rIIEαβ/WT, TFIIF, the Core-TFIIH, and the CAK, in presence of ATP (200 μM) and either XPD/WT, /R112H or /R722W. The immunoblot signals for TFIIEα, CDK7, and CyclinH were quantified (n = 3, means ± s.d.) and plotted in arbitrary units (a.u.). The results are representative of three independent experiments. c, d In vitro reconstituted transcription systems with rIIEs (c) and rIIHs (d) have been performed in presence of cold NTPs. Immunoblot analysis next has been done using specific antibody against Ser5-P of Pol II. Signals were quantified (n = 3, means ± s.d.) and plotted in arbitrary units (a.u.). The results are representative of three independent experiments. Source data are provided as a Source Data file
