title: Selective microRNA uridylation by Zcchc6 (TUT7) and Zcchc11 (TUT4).
abstract: Recent small RNA sequencing data has uncovered 3' end modification of mature microRNAs (miRNAs). This non-templated nucleotide addition can impact miRNA gene regulatory networks through the control of miRNA stability or by interfering with the repression of target mRNAs. The miRNA modifying enzymes responsible for this regulation remain largely uncharacterized. Here we describe the ability for two related terminal uridyl transferases (TUTases), Zcchc6 (TUT7) and Zcchc11 (TUT4), to 3' mono-uridylate a specific subset of miRNAs involved in cell differentiation and Homeobox (Hox) gene control. Zcchc6/11 selectively uridylates these miRNAs in vitro, and we biochemically define a bipartite sequence motif that is necessary and sufficient to confer Zcchc6/11 catalyzed uridylation. Depletion of these TUTases in cultured cells causes the selective loss of 3' mono-uridylation of many of the same miRNAs. Upon TUTase-dependent loss of uridylation, we observe a concomitant increase in non-templated 3' mono-adenylation. Furthermore, TUTase inhibition in Zebrafish embryos causes developmental defects and aberrant Hox expression. Our results uncover the molecular basis for selective miRNA mono-uridylation by Zcchc6/11, highlight the precise control of different 3' miRNA modifications in cells and have implications for miRNA and Hox gene regulation during development.
authors.affiliation: Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: James E
authors.initials: JE
authors.lastname: Thornton
authors.affiliation: Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Peng
authors.initials: P
authors.lastname: Du
authors.affiliation: Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Lili
authors.initials: L
authors.lastname: Jing
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Ljiljana
authors.initials: L
authors.lastname: Sjekloca
authors.affiliation: Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Shuibin
authors.initials: S
authors.lastname: Lin
authors.affiliation: Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Elena
authors.initials: E
authors.lastname: Grossi
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Piotr
authors.initials: P
authors.lastname: Sliz
authors.affiliation: Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA Harvard Stem Cell Institute, Cambridge, MA 02138, USA Department of Genetics, Harvard Medical School, Boston, MA 02115, USA Howard Hughes Medical Institute, Boston, MA 02115, USA.
authors.forename: Leonard I
authors.initials: LI
authors.lastname: Zon
authors.affiliation: Stem Cell Program, Boston Children's Hospital, Boston, MA 02115, USA Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA Harvard Stem Cell Institute, Cambridge, MA 02138, USA rgregory@enders.tch.harvard.edu.
authors.forename: Richard I
authors.initials: RI
authors.lastname: Gregory
mesh_terms: D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D005801:Genes, Homeobox; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry / Q000378:metabolism*; D059372:Nucleotide Motifs; D012316:RNA Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014529:Uridine / Q000378:metabolism*; D015027:Zebrafish / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D035683:MicroRNAs; C511130:TUT4 protein, human; D012316:RNA Nucleotidyltransferases; C576505:TUT7 protein, human; D014529:Uridine