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

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

Research output: Contribution to journalArticle

16 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

Fingerprint

Functional analysis
Double-Stranded RNA
Ribonuclease III
Missense Mutation
Small Interfering RNA
Drosophila
RNA-Induced Silencing Complex
Mutation
Processing
RNA Precursors
RNA Interference
Proteins
Genes
RNA

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Functional analysis of dicer-2 missense mutations in the siRNA pathway of Drosophila. / Lim, Do Hwan; Kim, Jung; Kim, Sanguk; Carthew, Richard W.; Lee, Youngsik.

In: Biochemical and Biophysical Research Communications, Vol. 371, No. 3, 04.07.2008, p. 525-530.

Research output: Contribution to journalArticle

Lim, Do Hwan ; Kim, Jung ; Kim, Sanguk ; Carthew, Richard W. ; Lee, Youngsik. / Functional analysis of dicer-2 missense mutations in the siRNA pathway of Drosophila. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 371, No. 3. pp. 525-530.
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