Fig. 2: Effects of TFIIF and its domains on activity of Pol II–Alu RNA complexes.
From: Mechanism of mammalian transcriptional repression by noncoding RNA
a, Fluorescence anisotropy experiments reveal Alu-LA and Alu-RA binding affinity to Pol II. Results represent analysis of three independently prepared reactions. Data are plotted as mean values ± standard deviation. b, Alu-LA density overlaid on Pol II-TFIIF as found within a transcription initiation open complex (PDB ID 5IYB)25. Pol II subunits are shown as semitransparent ribbons. The mutated regions of the TFIIF constructs used in c are indicated. c, Transcription assays carried out in the presence of the indicated TFIIF variant and either no RNA, Alu-LA or Alu-RA. Results represent analysis of three independently prepared reactions. Data are presented as mean values ± standard deviation, with individual data points shown as black circles. Achieved significance level relative to the wild-type TFIIF condition was calculated using an unpaired, two-tailed t-test: ***P ≤ 0.001; **P ≤ 0.01; *P ≤ 0.05; NS, P > 0.05. Exact P values are as follows: TFIIF (W164A), Alu-LA P = 0.0169, Alu-RA P = 0.2798; TFIIF (RAP30ΔC), Alu-LA P = 0.0002, Alu-RA P = 0.0040; TFIIF (RAP74ΔC), Alu-LA P = 0.0030, Alu-RA P = 0.0089. d, Schematic representation of the effects of TFIIF domains on the repressive activity of Alu-LA and Alu-RA.
