2021 Fiscal Year Research-status Report
Structural and molecular basis of the mitophagy regulation mediated by the Far complex in yeast
| Project/Area Number |
21K20632
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| Research Institution | Niigata University |
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Principal Investigator |
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| Project Period (FY) |
2021-08-30 – 2023-03-31
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| Keywords | Mitophagy / Atg32 / Ppg1 / The Far complex / Yeast / Autophagy |
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| Outline of Annual Research Achievements |
Mitophagy contributes to maintaining mitochondrial quality and quantity. The phosphorylation of Atg32 is essential for mitophagy. The Far complex interacts with Atg32 to phosphorylate it, but the underlying mechanism is still unclear. Therefore, this study aims to elucidate: 1) How the Far complex and Atg32 interact 2) the upstream signalling pathway regulating this interaction. So far I elucidated the next points: 1) None of genomically substituted Far8 phosphorylation sites mutants affected Atg32 phosphorylation status and Far8-Atg32 interaction. 2) I Found out that Far3/7-Atg32 interaction is mediated by Far8. Far3 and Far7 (parts of the Far complex) are necessary for Far8-Atg32 interaction.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
I had some problems with protein expression of plasmids for Far8 substitution mutants and I had to change strategy to genomic integration of Far8 substitution mutations, which takes longer time compared to construction of plasmids with substitution mutations
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| Strategy for Future Research Activity |
Next, I am planing to purify Atg32-Far complex for further Cryo-electron microspoic analysis of the bound state structure of Atg32-Far complex. Then, I am also planning to find out alternative posttranslational modification of Far8 and upstream singnaling pathway which regulate association and dissociation of Far8-Atg32 interaction.
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| Causes of Carryover |
Because of the delay caused by problems with Far8 substitution mutants plasmids, I had to move experiments with Atg32-Far complex purification to the next fiscal year.
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