Figure 10.

Identification of Can SINE regions required for polyadenylation of its pol III transcripts. (A) The Can sequence used in experiments (Can-T, above); the arrow indicates the transcription start; boxes A and B are underlined; polyadenylation signals (PASs) are shown in violet. Below is the alignment of the 3′-part of this sequence and derived constructs with modifications. The nucleotides different from the consensus in the region between the box B and TC-motif are given in red; blue marks the modifications in this region relative to the sequence above; modifications in the polypyrimidine region are shown in green. Deleted nucleotides, “-”. (B) Northern blot hybridization of RNA from HeLa cells transfected by Can constructs. The band and the smear above correspond to the primary and polyadenylated transcripts, respectively. Can-C is the construct with T-to-C substitutions in both polyadenylation signals, and its transcripts are not polyadenylated (For original image of Figure 10B, please refer to Supplementary Figure S11). (C) Polyadenylation efficiency of the modified constructs relative to Can-T. Error bars, SD, n = 3.