Lafora disease fibroblasts exemplify the molecular interdependence between thioredoxin 1 and the proteasome in mammalian cells

Abstract

Thioredoxin 1 (Trx1) is a key regulator of cellular redox balance and participates in cellular signaling events. Recent evidence from yeast indicates that members of the Trx family interact with the 20S proteasome, indicating redox regulation of proteasome activity. However, there is little information about the interrelationship of Trx proteins with the proteasome system in mammalian cells, especially in the nucleus. Here, we have investigated this relationship under various cellular conditions in mammalian cells. We show that Trx1 levels and its subcellular localization (cytosol, endoplasmic reticulum, and nucleus) depend on proteasome activity during the cell cycle in NIH3T3 fibroblasts and under stress conditions, when proteasomes are inhibited. In addition, we also studied in these cells how the main cellular antioxidant systems are stimulated when proteasome activity is inhibited. Finally, we describe a reduction in Trx1 levels in Lafora disease fibroblasts and demonstrate that the nuclear colocalization of Trx1 with 20S proteasomes in laforin-deficient cells is altered compared with control cells. Our results indicate a close relationship between Trx1 and the 20S nuclear proteasome and give a new perspective to the study of diseases or physiopathological conditions in which defects in the proteasome system are associated with oxidative stress.

Keywords: 78-kDa glucose-regulated protein; ATF4; Antioxidant enzymes; BIP/Grp78; BrdU; C/EBP-homologous protein (DNA damage-inducible transcript 3 protein); CHOP; Cell proliferation; CuZnSOD; DSB; ER; Free radicals; GSH; GSSG; Lafora disease; MnSOD; PARP; PDI; Proteasome; Rare diseases; Thioredoxin 1; Trx1; TrxR1; UPR; bromodeoxyuridine; copper–zinc superoxide dismutase; cyclic AMP-dependent transcription factor 4; double-strand break; endoplasmic reticulum; manganese superoxide dismutase; oxidized glutathione; poly(ADP-ribose) polymerase; protein disulfide isomerase; reduced glutathione; thioredoxin 1; thioredoxin reductase 1; unfolded protein response.