An RNAi-based genetic screen for oxidative stress resistance reveals retinol saturase as a mediator of stress resistance

Rie Nagaoka-Yasuda, Naoki Matsuo, Brian Perkins, Klara Limbaeck-Stokin, Mark Mayford

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Oxidative stress has been implicated in the pathogenesis of numerous late-onset diseases as well as organismal longevity. Nevertheless, the genetic components that affect cellular sensitivity to oxidative stress have not been explored extensively at the genome-wide level in mammals. Here we report an RNA interference (RNAi) screen for genes that increase resistance to an organic oxidant, tert-butylhydroperoxide (tert-BHP), in cultured fibroblasts. The loss-of-function screen allowed us to identify several short hairpin RNAs (shRNAs) that elevated the cellular resistance to tert-BHP. One of these shRNAs strongly protected cells from tert-BHP and H2O2 by specifically reducing the expression of retinol saturase, an enzyme that converts all-trans-retinol (vitamin A) to all-trans-13,14-dihydroretinol. The protective effect was well correlated with the reduction in mRNA level and was observed in both primary fibroblasts and NIH3T3 cells. The results suggest a novel role for retinol saturase in regulating sensitivity to oxidative stress and demonstrate the usefulness of large-scale RNAi screening for elucidating new molecular pathways involved in stress resistance.

Original languageEnglish
Pages (from-to)781-788
Number of pages8
JournalFree Radical Biology and Medicine
Volume43
Issue number5
DOIs
Publication statusPublished - Sep 1 2007
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Physiology (medical)

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