Fig. 1.

Putative miR-296-5p binding sites in PUMA mRNA. Schematic representation of the PUMA 3′UTR (#AF354654) nucleotides 863-1843. Computational algorithms were used to predict microRNAs that bind to a target sequence in the 3′UTR of PUMA. Two putative miR-296-5p binding sites were identified at position 1299/1306 (BS1) and position 1465/1471 (BS2) of PUMA 3′UTR. The complementarity of the 5′ seed region of miR-296-5p with BS1 and BS2 is indicated in the lower panel. According to Grimson et al. (17), BS1 is a 7mer site which contains the seed match (6 nucleotide at position 2-7) augmented by a match to microRNA nucleotide 8; BS2 is a 8mer site which contains the seed match flanked by both the match at position 8 and an A at position 1. As described in Materials and Methods, deletion or mutation of critical residues of BS1 and BS2 were generated for luciferase reporter gene assay.