| Budget Amount *help |
¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Outline of Final Research Achievements |
To identify novel GFP-LC3-interacting proteins in intestinal epithelial cells (IECs), we performed immunoprecipitation with a GFP antibody and then analyzed co- immunoprecipitates by mass spectrometry. HADHA was identified as an LC3-interacting protein. Given that HADHA catalyzes the last three steps of mitochondrial beta-oxidation of long-chain fatty acids, we investigated whether long-chain fatty acids induce autophagy. We found that palmitic acid- induced autophagy supports the survival of IECs.
Ubiquitination is frequently a prerequisite for substrate recognition and determines selectivity in autophagy. We developed a polyubiquitin-mediated fluorescence complementation (PolyUb-FC) assay. The PolyUb- FC assay has the advantage that chain-specific polyubiquitination can be directly visualized in living cells without using antibodies. We applied the PolyUb-FC assay to examine RIPK3 polyubiquitination. We demonstrated that RIPK3 colocalized with PolyUb-FC.
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