Cloning of the Oxidative Stress-responsive Genes in Caenorhabditis elegans

Abstract

Defense systems against free radicals and reactive oxygen species minimize the lethal and mutagenic consequences of these destructive agents. To investigate the genetic response to oxidative stress in a eukaryote, we cloned and characterized oxidative stress-responsive genes by comparing gene expression in the free-living nematode Caenorhabditis elegans under atmospheric conditions and high oxygen concentrations using a method of RNA arbitrarily primed polymerase chain reaction method (RAP-PCR). We identified four genes whose expression levels increase under high oxygen. These encoded 18s, 5.8 s and 26 s rRNAs, 16kDa heat shock proteins (hsp16–1 and 16–48) and a vacuolar ATPase G subunit. In addition, we also established that oxi-1, an oxidative stress-responsive gene we previously cloned, encodes a family of proteins related to human E6-AP ubiquitin-protein ligase. The similarity between human and nematode was 54% in one conserved amino acid region.