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First published online October 30, 2006
doi: 10.1242/10.1242/jcs.03229

Journal of Cell Science 119, 4554-4564 (2006)
Published by The Company of Biologists 2006
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Research Article

Glutaredoxins Grx3 and Grx4 regulate nuclear localisation of Aft1 and the oxidative stress response in Saccharomyces cerevisiae

Nuria Pujol-Carrion1, Gemma Belli1, Enrique Herrero1, Antoni Nogues2 and Maria Angeles de la Torre-Ruiz1,*

1 Departament de Ciències Mèdiques Bàsiques, Universitat de Lleida, Lleida 25198, Spain
2 Servei d'analisis cliniques, Hospital Arnau de Vilanova, Lleida 25192, Spain

* Author for correspondence (e-mail: madelatorre{at}cmb.udl.es)

Accepted 23 August 2006

Grx3 and Grx4, two monothiol glutaredoxins of Saccharomyces cerevisiae, regulate Aft1 nuclear localisation. We provide evidence of a negative regulation of Aft1 activity by Grx3 and Grx4. The Grx domain of both proteins played an important role in Aft1 translocation to the cytoplasm. This function was not, however, dependent on the availability of iron. Here we demonstrate that Grx3, Grx4 and Aft1 interact each other both in vivo and in vitro, which suggests the existence of a functional protein complex. Interestingly, each interaction occurred independently on the third member of the complex. The absence of both Grx3 and Grx4 induced a clear enrichment of G1 cells in asynchronous cultures, a slow growth phenotype, the accumulation of intracellular iron and a constitutive activation of the genes regulated by Aft1. The grx3grx4 double mutant was highly sensitive to the oxidising agents hydrogen peroxide and t-butylhydroperoxide but not to diamide. The phenotypes of the double mutant grx3grx4 characterised in this study were mainly mediated by the Aft1 function, suggesting that grx3grx4 could be a suitable cellular model for studying endogenous oxidative stress induced by deregulation of the iron homeostasis. However, our results also suggest that Grx3 and Grx4 might play additional roles in the oxidative stress response through proteins other than Aft1.

Key words: Grx3, Grx4, Oxidative stress, Iron homeostasis, Aft1, Cell cycle


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