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. 2009 Dec;15(12):2351–2363. doi: 10.1261/rna.1882009

FIGURE 4.

FIGURE 4.

Functions of MRE and PBE in poly(A) metabolism of cyclin B2 3′ UTR reporter RNAs. (A) Schematic representation of the cyclin B2 3′ UTR. (B) Kinetics of polyadenylation and deadenylation of wild-type RNA or substitution variants altering the MRE (MREmut) or PBE (sub61–88). 32P-labeled reporter RNAs were injected and assayed as in Figure 3B; samples were taken at 5, 6.5, 8, 10.5, 12.5, and ∼15 h p.f. AS and AL denote short and long poly(A) tail length, respectively (dotted lines). (C) Less extensive polyadenylation of the reporter RNA lacking the PBE. The pre-MBT samples (5 and 6.5 h p.f.) from the experiment shown in B were analyzed in the same gel, for direct comparison of the average poly(A) tail lengths (AAve, dotted lines). (D) Normal deadenylation kinetics of chimeric B2 3′ UTR reporter RNAs deleted of nucleotides 120–180 and the fused to hB4 3′ UTR sequences, which furnish CPE plus HEX functions. The wild-type (1–120 nt) and PBE or MRE substituted reporter RNAs were assayed as in B; samples were taken at 4.5, 6, 8, 9, 10.5, and 12 h p.f.