microRNA miR‐315‐5p regulates developmental growth in <i>Drosophila</i> wings by targeting <i>S6k</i>

Tissue growth in Drosophila is regulated by various factors, with microRNAs (miRNAs) emerging as key players over the past decade. However, the precise roles of miRNAs in growth regulation remain incompletely understood. In this study, we explored the biological role of miR-315 in wing growth regulation. Inhibition of miR-315-5p activity using a miR-315 sponge led to an increase in wing size, whereas its overexpression resulted in reduced wing size, primarily through a decrease in wing cell size. We identified ribosomal protein kinase p-70-S6k (S6k) as a target of miR-315-5p in relation to wing growth control. Overexpression of miR-315 reduced both total S6k and phosphorylated S6k protein levels in Drosophila S2 cells and wing discs. Additionally, a luciferase reporter assay confirmed that miR-315-5p directly binds to the 3'-untranslated region of S6k. Consistently, RNAi-mediated depletion of S6k led to smaller wings, primarily due to a reduction in cell size. Notably, co-overexpression of active S6k rescued the wing defects caused by miR-315 overexpression. Overall, these findings demonstrate that miR-315 regulates wing growth by suppressing S6k expression.