In the presence of aggregation-prone proteins, the cytosol and endoplasmic reticulum (ER) undergo a dramatic shift in their respective redox status, with the cytosol becoming more oxidized and the ER more reducing. However, whether and how changes in the cellular redox status may affect protein aggregation is unknown. Here, we show that C. elegans loss-of-function mutants for the glutathione reductase gsr-1 gene enhance the deleterious phenotypes of heterologous human, as well as endogenous worm aggregation-prone proteins. These effects are phenocopied by the GSH-depleting agent diethyl maleate. Additionally, gsr-1 mutants abolish the nuclear translocation of HLH-30/TFEB transcription factor, a key inducer of autophagy, and strongly impair the degradation of the autophagy substrate p62/SQST-1::GFP, revealing glutathione reductase may have a role in the clearance of protein aggregates by autophagy. Blocking autophagy in gsr-1 worms expressing aggregation-prone proteins results in strong synthetic developmental phenotypes and lethality, supporting the physiological importance of glutathione reductase in the regulation of misfolded protein clearance. Furthermore, impairing redox homeostasis in both yeast and mammalian cells induces toxicity phenotypes associated with protein aggregation. Together, our data reveal that glutathione redox homeostasis may be central to proteostasis maintenance through autophagy regulation.
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We thank CGC (Caenorhabiditis Genetics Center) and NBRP (National BioResource Project), Richard Morimoto, Christian Pohl, Jan Gruber, Guy Caldwell, Randy Blakely, Hong Zhang, Enriq Herrero, Anne Bertolotti, and Susan Lindquist for providing strains and plasmids and Leticia Lemus for help with yeast viability assays. Cristina Ayuso García and the Live Cell Imaging Facility, Karolinska Institutet, Sweden (supported by grants from the Knut and Alice Wallenberg Foundation, the Swedish Research Council, the Centre for Innovative Medicine and the Jonasson Centre at RIT, Sweden) are acknowledged for technical assistance. The Spanish Ministry of Economy and Competitiveness supported EF-S and VG (BFU2016–78265-P), PA (BFU2016–79313-P and MDM-2016–0687), and AM-V (BFU2015–64408-P). AM-V was also supported by the Instituto de Salud Carlos III (PI11/00072) and RPV-M (CPII16/00004, PI14/00949 and PI17/00011). All projects were cofinanced by the Fondo Social Europeo (FEDER). AM-V is a member of the GENIE and EU-ROS Cost Actions of the European Union and RPV-M is a Marie Curie Fellow (CIG322034, EU).
DG-G, JAM-L, and AM-V designed the study, performed most experiments using C. elegans, analyzed the data, and wrote the manuscript. BS-N and JC performed the embryo recordings, quantified the blebbing/exploding phenotypes, and contributed to the design of the study. FJN-G and FM-L carried out the initial candidate RNAi screen and generated several strains. CP-F, JG, and AC-M performed the mammalian cell experiments. CDL generated some strains and performed paralysis and DEM toxicity experiments. CN contributed essential reagents and analyzed the data. MDS and RPV-M generated C. elegans strains and carried out mechanosensory assays. EF-S, VG, RP, and EC contributed with the yeast experiments. PA performed the embryo confocal live imaging and immunostaining. All authors edited and revised the manuscript.
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Guerrero-Gómez, D., Mora-Lorca, J.A., Sáenz-Narciso, B. et al. Loss of glutathione redox homeostasis impairs proteostasis by inhibiting autophagy-dependent protein degradation. Cell Death Differ 26, 1545–1565 (2019). https://doi.org/10.1038/s41418-018-0270-9
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