Related Experiment Video
Updated: Feb 7, 2026

Activating Autophagy by Aerobic Exercise in Mice
Published on: February 3, 2017
An autophagy assay reveals the ESCRT-III component CHMP2A as a regulator of phagophore closure
Yoshinori Takahashi1, Haiyan He2, Zhenyuan Tang2
1Department of Pediatrics, Penn State College of Medicine, Hershey, PA, 17033, USA. ytakahashi@pennstatehealth.psu.edu.
Abstract:
The mechanism of phagophore closure remains unclear due to technical limitations in distinguishing unclosed and closed autophagosomal membranes. Here, we report the HaloTag-LC3 autophagosome completion assay that specifically detects phagophores, nascent autophagosomes, and mature autophagic structures. Using this assay, we identify the endosomal sorting complexes required for transport (ESCRT)-III component CHMP2A as a critical regulator of phagophore closure. During autophagy, CHMP2A translocates to the phagophore and regulates the separation of the inner and outer autophagosomal membranes to form double-membrane autophagosomes. Consistently, inhibition of the AAA-ATPase VPS4 activity impairs autophagosome completion. The ESCRT-mediated membrane abscission appears to be a critical step in forming functional autolysosomes by preventing mislocalization of lysosome-associated membrane glycoprotein 1 to the inner autophagosomal membrane. Collectively, our work reveals a function for the ESCRT machinery in the final step of autophagosome formation and provides a useful tool for quantitative analysis of autophagosome biogenesis and maturation.
More Related Videos
12:50Screening Assays to Characterize Novel Endothelial Regulators Involved in the Inflammatory Response
Published on: September 15, 2017
11:44A Primary Human Trophoblast Model to Study the Effect of Inflammation Associated with Maternal Obesity on Regulation of Autophagy in the Placenta
Published on: September 27, 2017
Related Concept Videos
Autophagy
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...
Components of Stress
Interestingly, the hidden cube faces also experience these stresses, equal and...
Components of Language
Angle Closure Glaucoma: Treatment
Epigenetic Regulation
GTPases and their Regulation
Large G-proteins,...