Figure 5.
Ccr4-Not Deadenylation Activity Is Reduced on Duplex Nucleic Acid Substrates
(A–C) Deadenylation by a recombinant Ccr4-Not complex of the 20-mer-A30 RNA substrate annealed to (A) a 20-nt DNA oligo complementary to the region upstream of the poly(A)30 tail; (B) a 30-nt DNA oligo; or (C) a 50-nt DNA oligo complementary to the entire 20-mer-A30 RNA. A control reaction performed on the 20-mer-A30 RNA in the same experiment is in Figures S6A and S6B.
(D) Deadenylation of an RNA substrate that contains an upstream polyuridine stretch that is predicted to form a stable hairpin-loop structure with the poly(A) tail. See also Figures S3D, S6C, and S6D.
M, molecular weight marker consisting of the 20-mer substrate with 0, 10, or 30 As. See also Figures S3 and S6.