A comprehensive analysis of precursor microRNA cleavage by human Dicer Academic Article uri icon

Overview

MeSH Major

  • DEAD-box RNA Helicases
  • MicroRNAs
  • RNA Cleavage
  • RNA Precursors
  • Ribonuclease III

abstract

  • Dicer cleaves double-stranded RNAs (dsRNAs) or precursor microRNAs (pre-miRNAs) to yield ≈ 22-nt RNA duplexes. The pre-miRNA structure requirement for human Dicer activity is incompletely understood. By large-scale in vitro dicing assays and mutagenesis studies, we showed that human Dicer cleaves most, although not all, of the 161 tested human pre-miRNAs efficiently. The stable association of RNAs with Dicer, as examined by gel shift assays, appears important but is not sufficient for cleavage. Human Dicer tolerates remarkable structural variation in its pre-miRNA substrates, although the dsRNA feature in the stem region and the 2-nt 3'-overhang structure in a pre-miRNA contribute to its binding and cleavage by Dicer, and a large terminal loop further enhances pre-miRNA cleavage. Dicer binding protects the terminal loop from digestion by S1 nuclease, suggesting that Dicer interacts directly with the terminal loop region.

publication date

  • November 2012

Research

keywords

  • Academic Article

Identity

Language

  • eng

PubMed Central ID

  • PMC3479397

Digital Object Identifier (DOI)

  • 10.1261/rna.033688.112

PubMed ID

  • 22984192

Additional Document Info

start page

  • 2083

end page

  • 92

volume

  • 18

number

  • 11