Fig. 5

BLV microRNA mediated cleavage of the AS1-L transcript. Shown on the X axis is the region of the BLV genome containing the BLV microRNAs (denoted by red rectangles). Graphed above is the percentage of high throughput sequencing reads showing evidence of cleavage at each base position. Libraries were prepared from total YR2 RNA and YR2 cells that were fractionated into cytoplasmic and nuclear enriched fractions. Below the X axis is shown the precise base pair position of cleavage for the peaks observed at miR-B1-3p and miR-B4-3p. The reference sequence is shown at the bottom and the individual reads above. The first 10 bp of the reads match the reference sequence, the sequence to the right of the cleavage point (arrow) corresponds to the RNA oligo ligated to the free 5′ end. Sites of cleavage were enriched between bp 10 and 11 of a subset of the mature microRNAs. The frequency of reads showing evidence of cleavage was increased in the cytoplasmic fraction. Free 5′ ends observed outside bp 10 and 11 of the microRNA in each sample are likely the product of RNA degradation