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N-ethylmaleimide sensitive factor (NSF) is a regulator of plasma membrane rupture and necrotic cell death

Murray, Emma
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Thesis/Dissertation
Date
2021
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Biomedical Sciences
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http://dx.doi.org/10.34944/dspace/6803
Abstract
Necrosis, originally considered chaotic, has been found to be regulated by distinct molecular pathways. To identify novel regulators of plasma membrane rupture, a hallmark of necrotic cell death, a genome-wide shRNA loss-of-function screen was performed. We identified SNARE complex members, mediators of canonical membrane fusion events, as necessary for Ca2+ and ROS-induced plasma membrane rupture andnecrosis. We targeted N-ethylmaleimide sensitive factor (NSF) due to its requirement in SNARE recycling, lack of gene homologs, and redox sensitivity. Deletion of Nsf protected against membrane rupture induced by various necrotic stimuli, yet did not influence apoptosis, suggesting specificity in programmed necrosis. We discovered that NSF localizes to sites of membrane blebbing and rupture. Additionally, SNARE-binding, ATPase activity, and redox modification are necessary for NSF’s role in necrosis. We generated conditional Nsf knockout mice and induced skeletal muscle injury via cardiotoxin injection. Loss of NSF protected against sarcolemmal rupture and myocyte death. Complementarily, cardiomyocyte-specific deletion of Nsf protected against ischemia-reperfusion injury. The SNARE complex regulator, NSF, is a key mediator of membrane rupture in necrotic cell death and a promising therapeutic target for numerous diseases.
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