Oxidative stress in Caenorhabditis elegans : protective effects of the Omega class glutathione transferase ( GSTO-1 ) Cora Burmeister * , Kai Lüersen * , Alexander Heinick * , Ayman Hussein † , Marzena Domagalski ‡ , Rolf D. Walter ‡ and Eva Liebau * ,1 * Institute of Animal Physiology, University of Muenster, Germany; † Faculty of Science, An-Najah National University, Nablus, Palestine; and ‡ Bernhard Nocht Institute, Hamburg, Germany 1 Correspondence: Institute for Animal Physiology, University of Muenster, Hindenburgplatz 55, Muenster 48143, Germany. E-mail: liebaue@uni-muenster.de To elucidate the function of Omega class glutathione transferases (GSTs) (EC 2.5.1.18) in multicellular organisms, the GSTO-1 from Caenorhabditis elegans ( GSTO-1 ; C29E4.7) was investigated. Disc diffusion assays using Escherichia coli overexpressing GSTO-1 provided a test of resistance to long-term exposure under oxidative stress. After affinity purification, the recombinant GSTO-1 had minimal catalytic activity toward classic GST substrates but displayed significant thiol oxidoreductase and dehydroascorbate reductase activity. Microinjection of the GSTO-1 -promoter green fluorescent protein construct and immunolocalization by electron microscopy localized the protein exclusively in the intestine of all postembryonic stages of C. elegans . Deletion analysis identified an ∼300-nucleotide sequence upstream of the ATG start site necessary for GSTO-1 expression. Site-specific mutagenesis of a GATA transcription factor binding motif in the minimal promoter led to the loss of reporter expression. Similarly, RNA interference (RNAi) of Elt-2 indicated the involvement of this gut-specific transcription factor in GSTO-1 expression. Transcriptional up-regulation under stress conditions of GSTO-1 was confirmed by analyzing promoter-reporter constructs in transgenic C. elegans strains. To investigate the function of GSTO-1 in vivo , transgenic animals overexpressing GSTO-1 were generated exhibiting an increased resistance to juglone-, paraquat-, and cumene hydroperoxide-induced oxidative stress. Specific silencing of the GSTO-1 by RNAi created worms with an increased sensitivity to several prooxidants, arsenite, and heat shock. We conclude that the stress-responsive GSTO-1 plays a key role in counteracting environmental stress.—Burmeister, C., Lüersen, K., Heinick, A., Hussein, A., Domagalski, M., Walter, R. D., Liebau, E. Oxidative stress in Caenorhabditis elegans : protective effects of the Omega class glutathione transferase ( GSTO-1 ). Key Words: antioxidant • RNA interference • GST
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Oxidative stress in Caenorhabditis elegans: protective effects of the Omega class glutathione transferase (GSTO-1)
Burmeister, Cora; Lüersen, Kai; Heinick, Alexander; Hussein, Ayman; Domagalski, Marzena; Walter, Rolf D.; Liebau, Eva
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, Volume 22 (2): 343
Fed of American Socs for Experimental Biology – Feb 1, 2008
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