Fig. 3: Positive patches on NELF-AC and the NELF-E RRM domain are both involved in eRNA-driven NELF dissociation.
From: Enhancer RNAs stimulate Pol II pause release by harnessing multivalent interactions to NELF
EMSAs carried out with PEC variants that comprise a the NELF-ABC-EΔRRM complex (NELF ΔRRM), or b a mutant NELF–ABCE complex, in which lysines and arginines situated in positively charged surface patches on NELF-C are mutated to uncharged residues (R291Q, K315M, K371M, K372M, K374M, K384M, K388M, R419Q, and R506Q) (NELF-C patch mutant)42, c Quantification of NELF release using data from a, b. Data for wild-type NELF (Fig. 2b) are shown to aid comparison. Binding curves were plotted and fitted as in Fig. 2e. d EMSA using a NELF double mutant (NELF patch mutant + NELF-E ΔRRM). The three NELF mutants utilized in these experiments were described previously42. All assays were performed as in Fig. 2b–d, using Arc eRNAs (1–50), (1–100), or (1–200). In e, f EMSAs of the NELF double mutant using Nr4a1-(a) and Nr4a1-(b) eRNA fragments are shown, respectively. Source data for (a–f) are provided in a Source Data file.
