Fig. 1.

Localization of a novel regulatory element in the Wee1 3′ UTR. (A) Schematic diagram showing the constructs used in the experiment. The GST coding region was fused to various Wee1 deletion 3′ UTRs, as indicated. The polyadenylation hexanucleotide is represented by a black hexagon; CPEs are represented by open circles; mutated CPEs are represented by “X”s; and the broken line indicates deletion of the 3′ UTR. In the 256–283 reporter construct, sequences 3′ of the Wee1 polyadenylation hexanucleotide were replaced by β-globin 3′ UTR sequence (dashed line). (B) Polyadenylation of the injected RNAs was assessed in time-matched immature (I) or progesterone-stimulated (P) oocytes by RNA ligation-coupled PCR using a forward primer specific to the GST coding region of the injected reporter mRNAs. Retarded mobility is indicative of polyadenylation. (C) Sequence and regulatory element composition of the wild-type and CPE-disrupted 256–283 region of the Wee1 3′ UTR. A candidate 17 nucleotide region (Wee17) responsible for progesterone-regulated polyadenylation is indicated.