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. Author manuscript; available in PMC: 2010 Jan 7.
Published in final edited form as: Mol Cell. 2009 Aug 27;35(6):868–880. doi: 10.1016/j.molcel.2009.08.004

Figure 7. GW182 Binding to PABP Is Required for miRNA-Mediated Deadenylation.

Figure 7

(A) Alignment of GW182 DUF sequences with Paip2 PAM2 motif.

(B) Western blot analysis of GST pulldowns of TNRC6C (1260-1690)-FLAG incubated with various C-terminal (C1 and C2) fragments of GST-PABP and probed with anti-FLAG and anti-GST antibodies.

(C) Western blot analysis of GST pulldowns of GST-PABP incubated with TNRC6C (1260-1690)-FLAG and/or wild-type or mutant PAM2 peptide and probed with anti-FLAG and anti-GST antibodies.

(D) 6xB-3′UTR RNA deadenylation in Krebs extract in the presence of increasing concentrations (1, 10, 50, and 100 μM) of wild-type or mutant (F > A) Paip2 PAM2 peptides.

(E) 6xB-3′UTR RNA deadenylation in Krebs extract, as determined by autoradiography, in the presence of increasing concentrations (0.5, 1.0, and 2.0 μg per reaction) of GST, TNRC6C (1260-1690), or TNRC6C (800-1360).