http://paymanhassibi.persiangig.com/document/mittler%20oxidative.pdf

Oxidative stress, antioxidants and stress tolerance

Antioxidants, Oxidative Damage and Oxygen Deprivation Stress: a Review

While the chemical nature of reactive oxygen species (ROS) dictates that they are potentially harmful to cells, recent genetic evidence suggests that in planta purely physicochemical damage may be much more limited than previously thought. The most potentially deleterious effect of ROS under most conditions is that at high concentrations they trigger genetically programmed cell suicide events. Moreover, because plants use ROS as second messengers in signal transduction cascades in processes as diverse as mitosis, tropisms and cell death, their accumulation is crucial to plant development as well as defence. Direct ROS signal transduction will ensue only if ROS escape destruction by antioxidants or are otherwise consumed in a ROS cascade. Thus, the major low molecular weight antioxidants determine the specificity of the signal. They are also themselves signal-transducing molecules that can either signal independently or further transmit ROS signals. The moment has come to re-evaluate the concept of oxidative stress. In contrast to this pejorative or negative term, implying a state to be avoided, we propose that the syndrome would be more usefully described as 'oxidative signalling', that is, an important and critical function associated with the mechanisms by which plant cells sense the environment and make appropriate adjustments to gene expression, metabolism and physiology.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-3040.2005.01327.x

Oxidant and antioxidant signalling in plants: a re-evaluation of the concept of oxidative stress in a physiological context

Increases in the intracellular levels of reactive oxygen species (ROS), frequently referred to as oxidative stress, represents a potentially toxic insult which if not counteracted will lead to membrane dysfunction, DNA damage and inactivation of proteins. Chronic oxidative stress has numerous pathological consequences including cancer, arthritis and neurodegenerative disease. Glutathione-associated metabolism is a major mechanism for cellular protection against agents which generate oxidative stress. It is becoming increasingly apparent that the glutathione tripeptide is central to a complex multifaceted detoxification system, where there is substantial inter-dependence between separate component members. Glutathione participates in detoxification at several different levels, and may scavenge free radicals, reduce peroxides or be conjugated with electrophilic compounds. Thus, glutathione provides the cell with multiple defences not only against ROS but also against their toxic products. This article discusses how glutathione biosynthesis, glutathione peroxidases, glutathione S-transferases and glutathione S-conjugate efflux pumps function in an integrated fashion to allow cellular adaption to oxidative stress. Co-ordination of this response is achieved, at least in part, through the antioxidant responsive element (ARE) which is found in the promoters of many of the genes that are inducible by oxidative and chemical stress. Transcriptional activation through this enhancer appears to be mediated by basic leucine zipper transcription factors such as Nrf and small Maf proteins. The nature of the intracellular sensor(s) for ROS and thiol-active chemicals which induce genes through the ARE is described. Gene activation through the ARE appears to account for the enhanced antioxidant and detoxification capacity of normal cells effected by many cancer chemopreventive agents. In certain instances it may also account for acquired resistance of tumours to cancer chemotherapeutic drugs. It is therefore clear that determining the mechanisms involved in regulation of ARE-driven gene expression has enormous medical implications.

Glutathione and glutathione-dependent enzymes represent a co-ordinately regulated defence against oxidative stress

The glutathione transferases (GSTs; also known as glutathione S-transferases) are major phase II detoxification enzymes found mainly in the cytosol. In addition to their role in catalysing the conjugation of electrophilic substrates to glutathione (GSH), these enzymes also carry out a range of other functions. They have peroxidase and isomerase activities, they can inhibit the Jun N-terminal kinase (thus protecting cells against H(2)O(2)-induced cell death), and they are able to bind non-catalytically a wide range of endogenous and exogenous ligands. Cytosolic GSTs of mammals have been particularly well characterized, and were originally classified into Alpha, Mu, Pi and Theta classes on the basis of a combination of criteria such as substrate/inhibitor specificity, primary and tertiary structure similarities and immunological identity. Non-mammalian GSTs have been much less well characterized, but have provided a disproportionately large number of three-dimensional structures, thus extending our structure-function knowledge of the superfamily as a whole. Moreover, several novel classes identified in non-mammalian species have been subsequently identified in mammals, sometimes carrying out functions not previously associated with GSTs. These studies have revealed that the GSTs comprise a widespread and highly versatile superfamily which show similarities to non-GST stress-related proteins. Independent classification systems have arisen for groups of organisms such as plants and insects. This review surveys the classification of GSTs in non-mammalian sources, such as bacteria, fungi, plants, insects and helminths, and attempts to relate them to the more mainstream classification system for mammalian enzymes. The implications of this classification with regard to the evolution of GSTs are discussed.

/pdf/structure-function-and-evolution-of-glutathione-transferases-10d1qis641.pdf

Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily.

Glutathione-mediated detoxification systems in plants.

Black-grass (Alopecurus myosuroides) is a major weed of wheat in Europe, with several populations having acquired resistance to multiple herbicides of differing modes of action. As compared with herbicide-susceptible black-grass, populations showing herbicide cross-resistance contained greatly elevated levels of a specific type I glutathione transferase (GST), termed AmGST2, but similar levels of a type III GST termed AmGST1. Following cloning and expression of the respective cDNAs, AmGST2 differed from AmGST1 in showing limited activity in detoxifying herbicides but high activities as a glutathione peroxidase (GPOX) capable of reducing organic hydroperoxides. In contrast to AmGST2, other GPOXs were not enhanced in the herbicide-resistant populations. Treatment with a range of herbicides used to control grass weeds in wheat resulted in increased levels of hydroperoxides in herbicide-susceptible populations but not in herbicide-resistant plants, consistent with AmGST2 functioning to prevent oxidative injury caused as a primary or secondary effect of herbicide action. Increased AmGST2 expression in black-grass was associated with partial tolerance to the peroxidizing herbicide paraquat. The selective enhancement of AmGST2 expression resulted from a constitutively high expression of the respective gene, which was activated in herbicide-susceptible black-grass in response to herbicide safeners, dehydration and chemical treatments imposing oxidative stress. Our results provide strong evidence that GSTs can contribute to resistance to multiple herbicides by playing a role in oxidative stress tolerance in addition to detoxifying herbicides by catalysing their conjugation with glutathione.

A role for glutathione transferases functioning as glutathione peroxidases in resistance to multiple herbicides in black-grass

Glutathione S-transferases (GSTs) from the phi (GSTF) and tau (GSTU) classes are unique to plants and play important roles in stress tolerance and secondary metabolism as well as catalyzing the detoxification of herbicides in crops and weeds. We have cloned and functionally characterized a group of GSTUs from wheat treated with fenchlorazole-ethyl, a herbicide safener. One of these enzymes, TaGSTU4-4, was highly active in conjugating the chemically distinct wheat herbicides fenoxaprop and dimethenamid. The structure of TaGSTU4-4 has been determined at 2.2 A resolution in complex with S-hexylglutathione. This enzyme is the first tau class GST structure to be determined and most closely resembles the omega class GSTs, but without the unique N-terminal extension or active site cysteine. The X-ray structure identifies key amino acid residues in the hydrophobic binding site and provides insights into the substrate specificity of these enzymes.

Structure of a tau class glutathione S-transferase from wheat active in herbicide detoxification.

Plants are able to metabolize agrochemicals and other foreign compounds by a variety of mechanisms and with extraordinary species diversity. Minor structural alterations of these compounds can bring about dramatic and unpredictable changes in the routes of their metabolism. The enzymes responsible for this exist in multiple forms which renders prediction of herbicide metabolism and, therefore, of selectivity, difficult. In some notable instances, pesticides are activated by plant metabolism. In the main, however, mechanisms such as hydroxylation, dealkylation and glutathione conjugation bring about detoxification and form the basis of herbicide selectivity. The properties of oxygenating and conjugating enzymes in plants are highlighted, with emphasis on the evident narrow substrate specificities, species differences and physiological roles. The molecular cloning of the genes specifying these enzymes will permit a much better definition of these mechanisms and will illuminate the natural roles of the enzymes involved. The prospects for utilizing recombinant enzymes as tools for the rational design of new selective herbicides are discussed. Herbicide safeners can protect certain crops from herbicide injury by promoting herbicide metabolism. The precise mechanisms of safener action and the reasons for their specificity are attracting much interest but are at present obscure. Natural variation in detoxification abilities of weed populations has allowed the field selection of some biotypes resistant to repeatedly used herbicides. By analogy, the introduction of microbial detoxification genes into major crops through genetic transformation has created new herbicideresistant crops which will enhance the flexibility of herbicide usage.

Detoxification and activation of agrochemicals in plants

The role of glutathione transferases (GSTs) in the selectivity of the herbicides alachlor, atrazine, fluorodifen and metolachlor, which are detoxified by glutathione conjugation in plants, was determined in seedlings of maize (Zea mays L.) and the associated weed species Abutilon theophrasti Medic., Digitaria sanguinalis (L.) Scop. Echinochloa crus-galli (L.) Beauv., Panicum miliaceum (L.), Setaria faberi Herrm. and Sorghum bicolor (L.) Moench. The availability of glutathione was also determined in all species and tissue concentrations were found to be in the range 120-160 μM for all species except D. sanguinalis and S. bicolor, which contained half this amount. GST activities toward the herbicides were determined in crude protein extracts from the plants using HPLC to quantify the biosynthesis of the herbicide conjugates. The specific activities of the GSTs toward the substrates were in the order alachlor > fluorodifen > atrazine > metolachlor in all species except A. theophrasti, where fluorodifen was a better substrate than alachlor. In most cases there was a good correlation between GST activities and the selectivity of the herbicides applied pre-emergence. In the case of atrazine, GST activities were also related to the relative rates of herbicide conjugation in vivo. In contrast, there was no simple relationship between glutathione availability and the selectivity of the herbicides. However, with alachlor there was evidence that glutathione availability was limiting GST activity and influencing tolerance.

David J. Cole

Papers

Glutathione-mediated detoxification systems in plants.

A role for glutathione transferases functioning as glutathione peroxidases in resistance to multiple herbicides in black-grass

Structure of a tau class glutathione S-transferase from wheat active in herbicide detoxification.

Detoxification and activation of agrochemicals in plants

Glutathione Transferase Activities and Herbicide Selectivity in Maize and Associated Weed Species