Characterization of Arabidopsis Mutants for the Variable Subunit of Ferredoxin:thioredoxin Reductase.
The ferredoxin/thioredoxin reductase (FTR) is the key enzyme of a light dependent redox regulatory system controlling enzyme activities in oxygenic photosynthetic cells. It is composed of two dissimilar subunits. The catalytic subunit contains a [4Fe-4S] cluster and a redox-active disulfide bridge as the active site. The function of the second subunit, named the variable subunit because it has less conserved primary sequence and length, is not yet known. In order to get insights into the physiological role and importance of FTR, we studied two Arabidopsis mutant lines in which one of two genes encoding FTRA subunit was disrupted by T-DNA insertion. In FTRA1 mutants, the absence of the corresponding transcript was not compensated by the increase in the level of FTRA2 mRNA. Mutant plants exhibited phenotypic perturbations when compared with wild-type plants. Disruptants were found significantly more sensitive to oxidative stress as imposed under high light or in the presence of paraquat. Mutants were further characterized at the biochemical level. Despite the fact that no difference was found by immunodetection of FTR polypeptides, evidence for an impaired FTR system occurring in the mutants was obtained by measuring the endogenous activation rate of one of its targets. In the leaves of mutants placed under normal culture conditions, NADP-dependent malate dehydrogenase (NADP-MDH) activation rate was abnormally low. A partially compensating increase of the enzyme activity was found as well as a higher amount of 2-cys-peroxiredoxin. Our results provide in planta confirmation of the antioxidant role previously proposed for some of the plastidial thioredoxins from Arabidopsis thaliana. The variable subunit of the FTR proved to be important, but its precise role remains to be established.
Keryer
Eliane
E
Institut de Biotechnologie des Plantes, UMR CNRS 8618, Bâtiment 630, Université Paris-Sud, 91405, Orsay Cedex, France.
Collin
Valérie
V
Lavergne
Danièle
D
Lemaire
Stéphane
S
Issakidis-Bourguet
Emmanuelle
E