Genome-wide identification of targets of the drosha-pasha/DGCR8 complex

RNA. 2009 Apr;15(4):537-45. doi: 10.1261/rna.1319309. Epub 2009 Feb 17.


Drosha is a type III RNase, which plays a critical role in miRNA biogenesis. Drosha and its double-stranded RNA-binding partner protein Pasha/DGCR8 likely recognize and cleave miRNA precursor RNAs or pri-miRNA hairpins cotranscriptionally. To identify RNAs processed by Drosha, we used tiling microarrays to examine transcripts after depletion of drosha mRNA with dsRNA in Drosophila Schneider S2 cells. This strategy identified 137 Drosha-regulated RNAs, including 11 putative pri-miRNAs comprising 15 annotated miRNAs. Most of the identified pri-miRNAs seem extremely large, >10 kb as revealed by both the Drosha knock-down strategy and by RNA PolII chromatin IP followed by Drosophila tiling microarrays. Surprisingly, more than a hundred additional RNAs not annotated as miRNAs are under Drosha control and are likely to be direct targets of Drosha action. This is because many of them encode annotated genes, and unlike bona fide pri-miRNAs, they are not affected by depletion of the miRNA processing factor, dicer-1. Moreover, application of the evofold analysis software indicates that at least 25 of the Drosha-regulated RNAs contain evolutionarily conserved hairpins similar to those recognized by the Drosha-Pasha/DGCR8 complex in pri-miRNAs. One of these hairpins is located in the 5' UTR of both pasha and mammalian DGCR8. These observations suggest that a negative feedback loop acting on pasha mRNA may regulate the miRNA-biogenesis pathway: i.e., excess Drosha cleaves pasha/DGCR8 primary transcripts and leads to a reduction in pasha/DGCR8 mRNA levels and Pasha/DGCR8 synthesis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 5' Untranslated Regions
  • Animals
  • Drosophila Proteins / chemistry
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / genetics*
  • Gene Knockdown Techniques
  • Genome, Insect*
  • Humans
  • MicroRNAs / genetics
  • Multiprotein Complexes / metabolism
  • Proteins / chemistry
  • Proteins / metabolism
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / metabolism*
  • Ribonuclease III / chemistry
  • Ribonuclease III / metabolism*


  • 5' Untranslated Regions
  • DGCR8 protein, human
  • Drosophila Proteins
  • MicroRNAs
  • Multiprotein Complexes
  • Pasha protein, Drosophila
  • Proteins
  • RNA-Binding Proteins
  • DROSHA protein, human
  • Ribonuclease III
  • drosha protein, Drosophila

Associated data

  • GEO/GSE14215