An RNAi-based genetic screen for oxidative stress resistance reveals retinol saturase as a mediator of stress resistance.
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 H(2)O(2) 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.
Nagaoka-Yasuda
Rie
R
Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Matsuo
Naoki
N
Perkins
Brian
B
Limbaeck-Stokin
Klara
K
Mayford
Mark
M
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D018384:Oxidative Stress; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
D012333:RNA, Messenger; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide