The Drosha-DGCR8 complex in primary microRNA processing
Top Cited Papers
Open Access
- 1 December 2004
- journal article
- research article
- Published by Cold Spring Harbor Laboratory in Genes & Development
- Vol. 18 (24), 3016-3027
- https://doi.org/10.1101/gad.1262504
Abstract
RNase III proteins play key roles in microRNA (miRNA) biogenesis. The nuclear RNase III Drosha cleaves primary miRNAs (pri-miRNAs) to release hairpin-shaped pre-miRNAs that are subsequently cut by the cytoplasmic RNase III Dicer to generate mature miRNAs. While Dicer (class III) and other simple RNase III proteins (class I) have been studied intensively, the class II enzyme Drosha remains to be characterized. Here we dissected the action mechanism of human Drosha by generating mutants and by characterizing its new interacting partner, DGCR8. The basic action mechanism of Drosha was found to be similar to that of human Dicer; the RNase III domains A and B form an intramolecular dimer and cleave the 3′ and 5′ strands of the stem, respectively. Human Drosha fractionates at ∼650 kDa, indicating that Drosha functions as a large complex. In this complex, Drosha interacts with DGCR8, which contains two double-stranded RNA (dsRNA)-binding domains. By RNAi and biochemical reconstitution, we show that DGCR8 may be an essential component of the pri-miRNA processing complex, along with Drosha. Based on these results, we propose a model for the action mechanism of class II RNase III proteins.Keywords
This publication has 34 references indexed in Scilit:
- Single Processing Center Models for Human Dicer and Bacterial RNase IIICell, 2004
- miRNAs on the move: miRNA biogenesis and the RNAi machineryCurrent Opinion in Cell Biology, 2004
- Identification of Virus-Encoded MicroRNAsScience, 2004
- MicroRNA precursors in motion: exportin-5 mediates their nuclear exportTrends in Cell Biology, 2004
- Distinct Roles for Drosophila Dicer-1 and Dicer-2 in the siRNA/miRNA Silencing PathwaysCell, 2004
- MicroRNAs: Genomics, Biogenesis, Mechanism, and FunctionCell, 2004
- Exportin 5 is a RanGTP-dependent dsRNA-binding protein that mediates nuclear export of pre-miRNAsRNA, 2004
- Nuclear Export of MicroRNA PrecursorsScience, 2004
- Structure and nucleic-acid binding of the Drosophila Argonaute 2 PAZ domainNature, 2003
- The crystal structure of the Argonaute2 PAZ domain reveals an RNA binding motif in RNAi effector complexesNature Structural & Molecular Biology, 2003