A single non-coding RNA locus produces small and long regulatory RNAs with distinct biological functions.

The impact of non-coding RNAs on stem cell biology and differentiation processes is an important and incompletely understood area of research. Using the testes of Drosophila melanogaster as a proven model for investigating these processes, we identified a polycistronic locus from which 2 non-coding transcripts, miR-317 and the long non-coding RNA Peony, are produced, with alternative polyadenylation implicated in regulation of their differential expression levels. We report here that each transcript has a distinct role in Drosophila testes; the absence of miR-317 leads to the emergence of germ-cell tumors in developing flies, and the increased expression of Peony results in the disruption of the muscle sheath covering the testis. We also show that miR-317 deficiency increases Notch signaling activity in somatic cyst cells, upregulates multiple predicted targets of miR-317, and drives germline tumorigenesis. Our findings establish miR-317 as a tumor suppressor controlling Notch signaling strength and uncover alternative polyadenylation as an unrecognized mechanism governing miRNA expression.