Functional analysis of dicer-2 missense mutations in the siRNA pathway of Drosophila

Do Hwan Lim, Jung Kim, Sanguk Kim, Richard W. Carthew, Young Sik Lee

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

The Drosophila RNase III enzyme Dicer-2 processes double-stranded RNA (dsRNA) precursors into small interfering RNAs (siRNAs). It also interacts with the siRNA product and R2D2 protein to facilitate the assembly of an RNA-induced silencing complex (RISC) that mediates RNA interference. Here, we characterized six independent missense mutations in the dicer-2 gene. Four mutations (P8S, L188F, R269W, and P365L) in the DExH helicase domain reduced dsRNA processing activity. Two mutations were located within an RNase III domain. P1496L caused a loss of dsRNA processing activity comparable to a null dicer-2 mutation. A1453T strongly reduced both dsRNA processing and RISC activity, and decreased the levels of Dicer-2 and R2D2 proteins, suggesting that this mutation destabilizes Dicer-2. We also found that the carboxyl-terminal region of R2D2 is essential for Dicer-2 binding. These results provide further insight into the structure-function relationship of Dicer, which plays a critical role in the siRNA pathway.

Original languageEnglish
Pages (from-to)525-530
Number of pages6
JournalBiochemical and biophysical research communications
Volume371
Issue number3
DOIs
Publication statusPublished - 2008 Jul 4

Keywords

  • Dicer-2
  • Drosophila
  • Mutation
  • R2D2
  • RNA interference
  • siRNA

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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