Roles of thioredoxin reductase during the aerobic life of Lactococcus lactis.
Thiol-disulfide bond balance is generally maintained in bacteria by thioredoxin reductase-thioredoxin and/or glutathione-glutaredoxin systems. Some gram-positive bacteria, including Lactococcus lactis, do not produce glutathione, and the thioredoxin system is presumed to be essential. We constructed an L. lactis trxB1 mutant. The mutant was obtained under anaerobic conditions in the presence of dithiothreitol (DTT). Unexpectedly, the trxB1 mutant was viable without DTT and under aerated static conditions, thus disproving the essentiality of this system. Aerobic growth of the trxB1 mutant did not require glutathione, also ruling out the need for this redox maintenance system. Proteomic analyses showed that known oxidative stress defense proteins are induced in the trxB1 mutant. Two additional effects of trxB1 were not previously reported in other bacteria: (i) induction of proteins involved in fatty acid or menaquinone biosynthesis, indicating that membrane synthesis is part of the cellular response to a redox imbalance, and (ii) alteration of the isoforms of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GapB). We determined that the two GapB isoforms in L. lactis differed by the oxidation state of catalytic-site cysteine C152. Unexpectedly, a decrease specific to the oxidized, inactive form was observed in the trxB1 mutant, possibly because of proteolysis of oxidized GapB. This study showed that thioredoxin reductase is not essential in L. lactis and that its inactivation triggers induction of several mechanisms acting at the membrane and metabolic levels. The existence of a novel redox function that compensates for trxB1 deficiency is suggested.
Vido
Karin
K
Unité de Recherches Laitières et Génétique Appliquée, INRA, Domaine de Vilvert, 78352 Jouy en Josas, France.
Diemer
Hélène
H
Van Dorsselaer
Alain
A
Leize
Emmanuelle
E
Juillard
Vincent
V
Gruss
Alexandra
A
Gaudu
Philippe
P
D000332:Aerobiosis; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D004229:Dithiothreitol; D015180:Electrophoresis, Gel, Two-Dimensional; D005798:Genes, Bacterial; D005978:Glutathione; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000096:biosynthesis; D007527:Isoenzymes / Q000096:biosynthesis; D013294:Lactococcus lactis / Q000201:enzymology / Q000502:physiology*; D009154:Mutation; D010084:Oxidation-Reduction; D020543:Proteome; D013880:Thioredoxin-Disulfide Reductase / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D007527:Isoenzymes; D020543:Proteome; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D013880:Thioredoxin-Disulfide Reductase; D005978:Glutathione; D004229:Dithiothreitol