Understanding the Role of Yeast Yme1 in Mitochondrial Function Using Biochemical and Proteomics Analyses
Abstract
:1. Introduction
2. Results
2.1. Effect on YME1 Deletion on Yeast Grow under Fermentative and Respiratory Conditions
2.2. Effects of YME1 Deletion on Mitochondrial Membrane Potential and Cardiolipin Content
2.3. Mitochondrial Proteomics Analysis of YME1 Deletion Reveals Defects in Mitochondrial Function under Fermentative Condition
2.4. Lost of YME1 Impair the Activities of Mitochondrial OXPHOS Complexes
3. Discussion
4. Materials and Methods
4.1. Yeast Strains
4.2. Media and Growth Conditions
4.3. Spot Tests
4.4. Cardiolipin Content and Membrane Potential Analyses
4.5. Yeast Chronological Life Span (CLS) Analysis
4.6. Mitochondria Isolation
4.7. Blue Native in-gel Respiratory Complex Activity Assays
4.8. The Complexes III and IV Activity in-solution Assays
4.9. Mass Spectrometry Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Mitochondrial Biological Process | Proteins Involved | FDR-Corrected p-Value |
---|---|---|
Mitochondrion-ER membrane tethering (GO:1990456) | Mdm10, Mdm12, Mmm1 | 2.4 × 10–2 |
Cardiolipin metabolic process (GO:0032048) | Ict1, Pgs1, Ups1, Ups2 | 2.0 × 10–2 |
Protein insertion into mitochondrial membrane (GO:0051204) | Mdm10, Mdm12, Mmm1, Tom70, Tom71, Sam37 | 2.0 × 10–3 |
Mitochondrial genome maintenance (GO:0000002) | Ilv5, Mdm10, Mdm12, Mdv1, Mgm1, Mip1, Mmm1, Rrg8, Tim17 | 1.4 × 10–4 |
Mitochondrial transmembrane transport (GO:1990542) | Fsf1, Mpc2, Mpc3, Tom70, Tom71, Xdj1, Tim23, Tim17, Tom6, Tim44 | 1.6 × 10–4 |
Mitochondrial Biological Process | Proteins Involved | FDR-Corrected p-Value |
---|---|---|
Aerobic electron transport chain (ETC) (GO:0019646) | Cor1, Cox13, Cox4, Cox5a, Cox6, Cox8, Cox9, Cyc1, Qcr10, Qcr2, Qcr7, Qcr8, Qcr9, Sdh1, Sdh2*, Sdh3, Sdh4, Sdh5 | 4.85 × 10–6 |
TCA cycle (GO:0006099) | Aco1, Cit1, Fum1, Idh1, Idh2, Kgd1, Mdh1, Mdh3, Sdh1, Sdh2, Sdh3, Sdh4, Sdh5 | 1.0 × 10–4 |
Mitochondrial ATP transmembrane transport (GO:1990544) | Aac1, Odc1, Odc2, Pet9 | 4.2 × 10–2 |
Fatty-acyl-CoA metabolic process (GO:0035337) | Faa1, Faa3, Faa4 | 4.2 × 10–2 |
Pyruvate metabolic process (GO:0006090) | Cdc19, Eno2, Glk1, Gpd1, Gpm1, Hxk1, Hxk2, Pdc1, Pfk1, Pfk2, Pgk1, Pyc1, Tdh3 | 5.6 × 10–4 |
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Kan, K.T.; Nelson, M.G.; Grant, C.M.; Hubbard, S.J.; Lu, H. Understanding the Role of Yeast Yme1 in Mitochondrial Function Using Biochemical and Proteomics Analyses. Int. J. Mol. Sci. 2022, 23, 13694. https://doi.org/10.3390/ijms232213694
Kan KT, Nelson MG, Grant CM, Hubbard SJ, Lu H. Understanding the Role of Yeast Yme1 in Mitochondrial Function Using Biochemical and Proteomics Analyses. International Journal of Molecular Sciences. 2022; 23(22):13694. https://doi.org/10.3390/ijms232213694
Chicago/Turabian StyleKan, Kwan Ting, Michael G. Nelson, Chris M. Grant, Simon J. Hubbard, and Hui Lu. 2022. "Understanding the Role of Yeast Yme1 in Mitochondrial Function Using Biochemical and Proteomics Analyses" International Journal of Molecular Sciences 23, no. 22: 13694. https://doi.org/10.3390/ijms232213694