Glutathione – a key factor of uranium tolerance in plant cells

Uranium (U) is a widespread occurring radioactive toxic heavy metal. It could be accumulated in plant roots and to a lesser extent in leaves. Hence, it is mandatory for plants to develop sophisticated tolerance strategies against this heavy metal.
Glutathion is one of the key players in this network, because of its ability to complex heavy metals, its redox-capacity, and/or as precursor in the biosynthesis of heavy metal-binding peptides, e.g. phytochelatines. The cytoplasmic glutathione content (reduced glutathione, GSH) dropped on the half in cell suspensions of canola (Brassica napus) within the first 30 minutes after U contact. However, the GSH recovery to the normal level was reached after 60 minutes. Various U concentrations (1 – 50 µM) caused different GSH kinetics indicating several defence reactions.
Artificial depletion of cytoplasmic GSH with 1-chloro-2,4-dinitrobenzene (CDNB) enhances the U toxicity in cells determined by a metabolic test.
Because of a clear excess of GSH against the slowly accumulating U in the cytoplasm, the massive GSH decrease could not exclusively be dedicated to a complex formation between GSH and U. Additionally, a reduction of U(VI) to U(IV) can be ruled out because of a lack of U(IV) in cytoplasm revealed by photoacoustic measurements. However, the rapid GSH drop could be caused by its oxidation to GSSG (oxidized GSH). One possibility is the involvement of GSH in detoxification mechanisms against oxidative burst, e.g. reactive oxygen species (ROS), induced by U. Another could be the GSH dependent recovery of redox equivalents or other metabolites. Consequently, this GSH based detoxification processes will, at least transiently, generate a redox signal and therefore impact on cellular redox poise.
It has to figure out weather the direct complexation of U with GSH (see abstract Geipel et al.) or the conjugation via glutathione S-transferase and the subsequent transport to the vacuole takes place. However, the formation of phytochelatines cannot be excluded because of cytoplasmic U bounded proteins being smaller than 14 KDa revealed by SDS gel electrophoresis and subsequent fluorescence measurements.
Further investigations should provide more detailed insights in the intracellular network of GSH functions in U detoxification as a redox buffer and as a detoxification reagent.