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FBS Colloquia No.280Laboratory of Mitochondrial Dynamics

Seminar or Lecture

Research on the mitophagy initiation complex using an in vivo protein-protein interaction assay

YANG LIU [Graduate student (D5/5)・JSPS Research Fellow DC1, Laboratory of Mitochondrial Dynamics]

Date and Time 7 Oct. 2021 (Thu), 12:15~13:00
Place Online (Zoom) | An email will be sent with the meeting URL, ID, and password to all FBS members.
Language English
Contact

Koji Okamoto, Associate Professor
Tel: 06-6879-7970
E-mail: kokamoto[at]fbs.osaka-u.ac.jp

Research on the mitophagy initiation complex using an in vivo protein-protein interaction assay

Mitochondria-specific autophagy, termed mitophagy, is a fundamental catabolic pathway that mediates selective degradation of defective or excess mitochondria, contributing to mitochondrial quality and quantity control and cellular homeostasis. Mitophagy is an intracellular process conversed from yeast to humans, and impairments in mitophagy are thought to be associated with various disorders such as neurodegeneration, heart/liver failure, aging, and cancer. In budding yeast, the single-pass membrane protein Atg32 accumulates on the surface of mitochondria and forms a mitophagy initiation complex via its interactions with Atg11, a scaffold promoting assembly of other Atg proteins required for autophagosome formation, and Atg8, a ubiquitin-like fold that is conjugated to the phospholipid phosphatidylethanolamine and mediates autophagosome formation. Although this complex is crucial for mitophagy, how its formation is regulated and how it functions in the initiation events remain poorly understood. To solve these issues, we have applied NanoLuc Binary Technology (NanoBiT) to establish a quantitative assay for Atg32-Atg11 and Atg32-Atg8 interactions. This assay is a bioluminescence-based detection system capable of analyzing protein-protein interactions in live cells that is smaller-scale and faster compared to co-immunoprecipitation. In this colloquium, we will summarize our latest data on proteins critical for Atg32-Atg11 and Atg32-Atg8 interactions and discuss about how the mitophagy initiation complex drives formation of autophagosomes surrounding mitochondria.

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