Fig. 4

A peptide motif from Drosophila Bag-of-marbles inhibits deadenylation. a EMSA with the 7-mer-A₂₀ RNA and the CAF40 module in the absence or presence of MBP-tagged Bam CBM (wildtype or the double mutant). Only Bam CBM^WT, but not Bam CBM^MUT efficiently competed with RNA for CAF40 binding. b Titration experiment with 50 nM of the 7-mer-A₂₀ RNA and the CCR4-NOT_CORE complex, and the indicated concentrations (0.05–5.0 µM) of either MBP, which served as a negative control, or MBP-tagged Bam CBM^WT. A 10-fold excess (500 nM) of Bam CBM^WT was sufficient to almost completely inhibit deadenylation. The reactions were stopped after 32 min. c Time course assay with the 7-mer-A₂₀ RNA and the CCR4-NOT_CORE complex in equimolar ratio (50 nM). In addition, a 50-fold molar excess (2.5 µM) of either MBP, which served as a negative control, or MBP-tagged Bam CBM^WT or Bam CBM^MUT (containing L17E and M24E mutations) were included. d–f Titration experiments with 50 nM of the 7-mer-A₂₀ RNA and the CCR4-NOT_MINI (d), CCR4-NOT_FULL (e) or CCR4-NOT_Δ10:11 (f) complexes, respectively, and the indicated concentrations (0.05–5.0 µM) of either MBP, which served as a negative control, or MBP-tagged Bam CBM^WT. The reactions were stopped after 32 min. Source data are provided as a Source Data file