Figure 6.

PABP release requires the interaction of GW182 proteins with the CCR4–NOT complex. (A, B) The interactions of the TNRC6C-SD protein (wild-type or mutant) with NOT1 and PABP were analysed by coimmunoprecipitation using anti-GFP antibodies. GFP-tagged F-Luc served as a negative control. (C–F) S2 cells were transfected with a mixture of three plasmids as described in Figure 5. (C) Firefly luciferase activity was normalized to that of the Renilla luciferase and set at 100 in cells expressing the λN-HA peptide (white bar). (D) Northern blot of representative RNA samples analysed as described in Figure 1. (E) The association of the F-Luc-5BoxB-(A)₉₅-HhR reporter with the endogenous PABP in the absence (white bars) or presence (blue bars) of TNRC6C-SD was analysed as described in Figure 1. (F) Western blot showing comparable expression of TNRC6C-SD wild-type and mutants.

Source data for this figure is available on the online supplementary information page.

Source Data for Figure 6 ^{(4.5MB, pdf)}