title: The 3'-5' exoribonuclease Dis3 regulates the expression of specific microRNAs in Drosophila wing imaginal discs.
abstract: Dis3 is a highly conserved exoribonuclease which degrades RNAs in the 3'-5' direction. Mutations in Dis3 are associated with a number of human cancers including multiple myeloma and acute myeloid leukemia. In this work, we have assessed the effect of a Dis3 knockdown on Drosophila imaginal disc development and on expression of mature microRNAs. We find that Dis3 knockdown severely disrupts the development of wing imaginal discs in that the flies have a "no wing" phenotype. Use of RNA-seq to quantify the effect of Dis3 knockdown on microRNA expression shows that Dis3 normally regulates a small subset of microRNAs, with only 11 (10.1%) increasing in level ≥ 2-fold and 6 (5.5%) decreasing in level ≥ 2-fold. Of these microRNAs, miR-252-5p is increased 2.1-fold in Dis3-depleted cells compared to controls while the level of the miR-252 precursor is unchanged, suggesting that Dis3 can act in the cytoplasm to specifically degrade this mature miRNA. Furthermore, our experiments suggest that Dis3 normally interacts with the exosomal subunit Rrp40 in the cytoplasm to target miR-252-5p for degradation during normal wing development. Another microRNA, miR-982-5p, is expressed at lower levels in Dis3 knockdown cells, while the miR-982 precursor remains unchanged, indicating that Dis3 is involved in its processing. Our study therefore reveals an unexpected specificity for this ribonuclease toward microRNA regulation, which is likely to be conserved in other eukaryotes and may be relevant to understanding its role in human disease.
authors.affiliation: a Brighton and Sussex Medical School; Medical Research Building; University of Sussex; Falmer , Brighton , UK.
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Towler
authors.forename: Christopher I
authors.initials: CI
authors.lastname: Jones
authors.forename: Sandra C
authors.initials: SC
authors.lastname: Viegas
authors.forename: Patricia
authors.initials: P
authors.lastname: Apura
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Waldron
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Smalley
authors.forename: Cecilia M
authors.initials: CM
authors.lastname: Arraiano
authors.forename: Sarah F
authors.initials: SF
authors.lastname: Newbury
mesh_terms: D000818:Animals; D004330:Drosophila / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism*; D005095:Exoribonucleases / Q000378:metabolism; D063326:Exosome Multienzyme Ribonuclease Complex / Q000235:genetics* / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D060227:Imaginal Discs / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D059467:Transcriptome; D014231:Tribolium; D014921:Wings, Animal / Q000254:growth & development / Q000378:metabolism
chemical_list: C502521:Dis3 protein, Drosophila; D029721:Drosophila Proteins; D035683:MicroRNAs; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex
keywords: Dis3/Taz; Drosophila development; RNA degradation; RNA stability; exoribonuclease; imaginal discs; miRNAs