The H/ACA complex disrupts triplex in hTR precursor to permit processing by RRP6 and PARN

Human telomerase RNA (hTR) is transcribed as a precursor that is then posttranscriptionally modified and processed. A fraction of the transcripts is oligoadenylated by TRAMP and either processed into the mature hTR or degraded by the exosome. Here, we characterize the processing of 3’ extended forms of varying length by PARN and RRP6. We show that tertiary RNA interactions unique to the longer precursor forms favor RNA degradation, whereas H/ACA RNP assembly stimulates productive processing. Interestingly, the H/ACA complex actively promotes processing in addition to protecting the mature 3’ end. Processing occurs in two steps with longer forms being trimmed by RRP6 and shorter forms being preferentially processed by PARN. These results reveal how RNA structure and RNP assembly affect the kinetics of processing and degradation and ultimately determine the amount of functional telomerase produced in cells. We cloned and sequenced 3’ ends by 3' RACE coupled with high-throughput sequencing to gain further insights into hTR processing.