The mitochondrial thioredoxin system regulates the TCA cycle-derived metabolic fluxes toward the GS/GOGAT cycle in illuminated leaves

Abstract

Previous studies suggest that the synthesis of glutamate/glutamine is regulated by the mitochondrial thioredoxin (TRX) system. However, the mechanisms behind it remains unclear. Here, we demonstrated that the level of citrate and glutamate was higher in illuminated leaves from Arabidopsis mutants lacking the mitochondrial TRX o1 (trxo1) or both NADPH-dependent TRX reductases A/B (ntrab), that are found in nucleus, cytosol and mitochondria, when compared to the wild type (WT). Increased ¹³C-labelling in glutamate derived from ¹³C-pyruvate was observed in illuminated trxo1 and ntrab leaves, but not in WT and in the microsomal trxh2 mutant. The lack of TRX o1 decreased the content and activity of glutamine synthetase (GS), which leads to a lower level of glutamine, and exacerbated the increases in GS activity triggered by high light, when compared to the WT. The level of glutamine was positively correlated with the percentage of the oxidized GS band. However, the GS redox status was unaltered in all mutants. Our results indicate mitochondrial TRX mutants have higher metabolic fluxes from the TCA to the GS/GOGAT cycle in vivo, likely associated to an increased substrate availability and by direct-and-indirect TRX-mediated mechanisms that regulate enzymes of both TCA and GS/GOGAT cycles.

Glutamate, glutamine, high light, redox regulation, thioredoxin reductases

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