title: RNase activity requires formation of disulfide bonds and is regulated by the redox state.
abstract: The activity of many RNases requires the formation of one or more disulfide bonds which can contribute to their stability. In this study, we show that RNase activity and, to a much lesser extent, nuclease activity, are redox regulated. Intracellular RNase activity was altered in vitro by changes in the glutathione redox state. Moreover, RNase activity was abolished following exposure to reducing agents such as beta-ME or DTT. Following reduction with glutathione (GSH), RNase activity could be fully reactivated with oxidized glutathione (GSSG). In contrast, RNase activity could not be reactivated when reduced with DTT. Decreasing the level of glutathione in vivo in wheat increased RNase activity. Tobacco engineered to have an increased glutathione redox state exhibited substantially lower RNase activity during dark-induced senescence. These results suggest that RNase activity requires the presence of one or more disulfide bonds that are regulated by glutathione and demonstrate for the first time that RNase activity can be altered with an alteration in cellular redox state.
authors.affiliation: Department of Biochemistry, University of California, CA, USA.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Chen
authors.forename: Jun
authors.initials: J
authors.lastname: Ling
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Gallie
mesh_terms: D003624:Darkness; D003851:Deoxyribonucleases / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects / Q000528:radiation effects; D005978:Glutathione / Q000378:metabolism / Q000494:pharmacology; D019803:Glutathione Disulfide / Q000378:metabolism / Q000494:pharmacology; D049691:History, Medieval; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D012260:Ribonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014908:Triticum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
chemical_list: D004220:Disulfides; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; C020666:glutathione dehydrogenase (ascorbate); D003851:Deoxyribonucleases; D012260:Ribonucleases; D005978:Glutathione; D019803:Glutathione Disulfide