2017 Fiscal Year Final Research Report
Analysis of negative regulator of necroptosis
Project/Area Number |
16K15423
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Research Category |
Grant-in-Aid for Challenging Exploratory Research
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Allocation Type | Multi-year Fund |
Research Field |
Gastroenterology
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Research Institution | Tokyo Medical and Dental University |
Principal Investigator |
OSHIMA Shigeru 東京医科歯科大学, 医学部附属病院, 助教 (50376787)
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Project Period (FY) |
2016-04-01 – 2018-03-31
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Keywords | オートファジー / ユビキチン / ネクロプトーシス |
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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Free Research Field |
消化器内科
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