Yeasts as Complementary Model Systems for the Study of the Pathological Repercussions of Enhanced Synphilin-1 Glycation and Oxidation
Abstract
:1. Introduction
2. Results
2.1. SY1 Production Results in a Growth Defect of S. cerevisiae Strains Devoid of Glo2 and Gre3
2.2. Disrupting Methylglyoxalase System and Aldose Reductase Activity Increases the Number of Large dsRed-SY1 Inclusions in S. cerevisiae
2.3. Immunofluorescent Detection of SY1 in S. pombe Reveals Polar-Localised Inclusions
2.4. Disrupting Methylglyoxalase System and Aldose Reductase Activity Is Associated with Reduced SY1 Protein Content in S. cerevisiae
2.5. Potential Localisation of dsRed-SY1 Inclusions to IPOD in Methylglyoxalase System and Aldose Reductase-Deficient Strains Indicates the Involvement of Autophagy
2.6. Disrupting Methylglyoxalase System and Aldose Reductase Activity Is Associated with Enhanced Oxidative Stress Levels in S. cerevisiae
2.7. Analysis of Actin Cytoskeleton Organisation in S. cerevisiae Cells Producing dsRed-SY1
3. Discussion
4. Materials and Methods
4.1. Cell Culture, Strains, and Plasmids
4.2. Growth Analyses and Spot Assays
4.3. Oxidative Stress Measurements
4.4. Fluorescence Microscopic Analysis
4.5. Immunological Techniques
4.5.1. Western Blot Analysis
4.5.2. Immunofluorescence Microscopy
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
PD | Parkinson’s disease |
αSyn | α-Synuclein |
SY1 | Synphilin-1 |
AGE | advanced glycation end-products |
ROS | reactive oxygen species |
MGO | Methylglyoxal OD optical density |
EV | empty vector |
ns | not significant |
CytoQ | cytoplasmic quality control compartment |
JUNQ/INQ | juxta/intranuclear quality control compartment |
IPOD | insoluble protein deposit |
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Glo1 | Glo2 | Gre3 | ||
---|---|---|---|---|
S. cerevisiae | gene | GLO1 (Lactoylglutathione lyase) | GLO2 and GLO4 (Hydroxyacylglutathione hydrolase) | GRE3 (NADPH-dependent aldose reductase) |
function | catalyzes the formation of S-D-lactoylglutathione | catalyzes the hydrolysis of S-D-lactoyl-glutathione to form glutathione and D-lactic acid | reduces methylglyoxal to acetol and (R)-lactaldehyde | |
S. pombe Ortholog | gene | glo1+ (Lactoylglutathione lyase) | glo2+ (Hydroxyacylglutathione hydrolase) | non existing |
function | the same as S. cerevisiae Glo1 | function not experimentally validated yet | no S. pombe aldose reductase has reported MGO-detoxifying activity | |
Human Ortholog | gene | GLO1 (Lactoylglutathione lyase) | GLO2 (Hydroxyacylglutathione hydrolase) | AKR1B10 (aldo-keto reductase family 1 member B10) |
function | the same as S. cerevisiae Glo1 | the same as S. cerevisiae Glo2 | catalyzes the NADPH-dependent reduction of a wide variety of carbonyl-containing compounds |
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Seynnaeve, D.; Mulvihill, D.P.; Winderickx, J.; Franssens, V. Yeasts as Complementary Model Systems for the Study of the Pathological Repercussions of Enhanced Synphilin-1 Glycation and Oxidation. Int. J. Mol. Sci. 2021, 22, 1677. https://doi.org/10.3390/ijms22041677
Seynnaeve D, Mulvihill DP, Winderickx J, Franssens V. Yeasts as Complementary Model Systems for the Study of the Pathological Repercussions of Enhanced Synphilin-1 Glycation and Oxidation. International Journal of Molecular Sciences. 2021; 22(4):1677. https://doi.org/10.3390/ijms22041677
Chicago/Turabian StyleSeynnaeve, David, Daniel P. Mulvihill, Joris Winderickx, and Vanessa Franssens. 2021. "Yeasts as Complementary Model Systems for the Study of the Pathological Repercussions of Enhanced Synphilin-1 Glycation and Oxidation" International Journal of Molecular Sciences 22, no. 4: 1677. https://doi.org/10.3390/ijms22041677