Lin28 Recruits the TUTase Zcchc11 to Inhibit let-7 Maturation in Mouse Embryonic Stem Cells

Nat Struct Mol Biol. 2009 Oct;16(10):1021-5. doi: 10.1038/nsmb.1676. Epub 2009 Aug 27.

Abstract

Lin28 and Lin28B, two developmentally regulated RNA-binding proteins and likely proto-oncogenes, selectively inhibit the maturation of let-7 family microRNAs (miRNAs) in embryonic stem cells and certain cancer cell lines. Moreover, let-7 precursors (pre-let-7) were previously found to be terminally uridylated in a Lin28-dependent fashion. Here we identify Zcchc11 (zinc finger, CCHC domain containing 11) as the 3' terminal uridylyl transferase (TUTase) responsible for Lin28-mediated pre-let-7 uridylation and subsequent blockade of let-7 processing in mouse embryonic stem cells. We demonstrate that Zcchc11 activity is UTP-dependent, selective for let-7 and recruited by Lin28. Furthermore, knockdown of either Zcchc11 or Lin28, or overexpression of a catalytically inactive TUTase, relieves the selective inhibition of let-7 processing and leads to the accumulation of mature let-7 miRNAs and repression of let-7 target reporter genes. Our results establish a role for Zcchc11-catalyzed pre-let-7 uridylation in the control of miRNA biogenesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Catalysis
  • Cell Line, Tumor
  • DNA-Binding Proteins / metabolism*
  • Embryonic Stem Cells / metabolism
  • Genes, Reporter
  • Mice
  • MicroRNAs / metabolism*
  • Models, Genetic
  • Oocytes / metabolism
  • Protein Structure, Tertiary
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / metabolism
  • RNA-Binding Proteins / physiology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Zinc Fingers

Substances

  • DNA-Binding Proteins
  • Lin-28 protein, mouse
  • MicroRNAs
  • RNA, Messenger
  • RNA-Binding Proteins
  • mirnlet7 microRNA, mouse
  • ZCCHC11 protein, mouse