The budding yeast orthologue of Parkinson's disease-associated DJ-1 is a multi-stress response protein protecting cells against toxic glycolytic products

Abstract

Saccharomyces cerevisiae Hsp31p is a DJ-1/ThiJ/PfpI family protein that was previously shown to be important for survival in the stationary phase of growth and under oxidative stress. Recently, it was identified as a chaperone or as glutathione-independent glyoxalase. To elucidate the role played by this protein in budding yeast cells, we investigated its involvement in the protection against diverse environmental stresses. Our study revealed that HSP31 gene expression is controlled by multiple transcription factors, including Yap1p, Cad1p, Msn2p, Msn4p, Haa1p and Hsf1p. These transcription factors mediate the HSP31 promoter responses to oxidative, osmotic and thermal stresses, to potentially toxic products of glycolysis, such as methylglyoxal and acetic acid, and to the diauxic shift. We also demonstrated that the absence of the HSP31 gene sensitizes cells to these stressors. Overproduction of Hsp31p and its homologue Hsp32p rescued the sensitivity of glo1Δ cells to methylglyoxal. Hsp31p also reversed the increased sensitivity of the ald6Δ strain to acetic acid. Since Hsp31p glyoxalase III coexists in S. cerevisiae cells with thousand-fold more potent glyoxalase I/II system, its biological purpose requires substantiation. We postulate that S. cerevisiae Hsp31p may have broader substrate specificity than previously proposed and is able to eliminate various toxic products of glycolysis. Alternatively, Hsp31p might be effective under high concentration of exogenous methylglyoxal present in some natural environmental niches populated by budding yeast, when glyoxalase I/II system capacity is saturated.

Oxidative stress triggers aggregation of GFP-tagged Hsp31p, the budding yeast environmental stress response chaperone, and glyoxalase III

“…This subcellular localization pattern is common to hundreds of S. cerevisiae proteins but does not provide any useful information regarding the intracellular role of Hsp31p. Nevertheless, we have previously demonstrated that Hsp31p is a multi-stress response protein (Natkańska et al 2017 ; Skoneczna et al 2007 ) that is responsive to the diauxic shift and to oxidative, osmotic, and thermal stresses. Moreover, strongest expression of the HSP31 gene was observed following exposure of yeast cells to toxic products of fermentation (ethanol and methylglyoxal).…”

“…Single-copy plasmids encoding various GFP-tagged Hsp31p proteins were constructed in several steps. To construct the single-copy YCpHSP31C➔A plasmid, the Eco RI- Nae I fragment containing a full-length mutated HSP31 gene (encoding a protein with a Cys138-to-Ala substitution), together with its promoter (up to −453 bp) and terminator (down to +277 bp), was excised from pRSHSPC➔A (Natkańska et al 2017 ) and cloned into a YCp50 centromeric vector (Rose et al 1987 ), linearized with the Eco RI and Nru I enzymes. To construct the YCpHSP31 plasmid containing the native HSP31 gene, the Hind III- Sac I insert was excised from pRSHSP31 (Natkańska et al 2017 ) and cloned into the YCpHSP31C➔A plasmid, linearized with the same enzymes.…”

“…Redox Western blot analysis of Hsp31p was performed with cytosolic rxYFP (a redox-sensitive protein) as the control according to a previously described method (Østergaard et al 2004 ). Briefly, the BY hsp31 ∆ strain was transformed with the pJH208 plasmid encoding the redox-sensitive rxYFP control protein (Hu et al 2008 ) and either the YEpHSP31 or YEpHSPC➔A plasmid (Natkańska et al 2017 ) encoding native Hsp31p or Hsp31p lacking Cys138, respectively. The cells were grown overnight in selective SC medium without uracil and leucine.…”

“…DJ-1 is by far the most frequently studied protein in this family. Nevertheless, the exact roles of DJ-1 in human cells (Chan and Chan 2015 ), Candida albicans Glx3, which belongs to the same family (Hasim et al 2014 ), and S. cerevisiae Hsp31p (Natkańska et al 2017 ; Aslam and Hazbun 2016 ) are not fully understood. One helpful clue for deciphering their biological role(s) is their cellular localization.…”

Metals and Parkinson's Disease: Mechanisms and Biochemical Processes

“…Interestingly, a DJ-1 protein that is known as a PD-associated protein protects cells from toxic stresses and can bind both mercury and lead [102]. Genetic variants of DJ-1 protein exert no protective effects on mercury toxicity and, therefore, increase the risk for PD [102,103].…”