Cholesterol transfer via endoplasmic reticulum contacts mediates lysosome damage repair
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Lysosome integrity is essential for cell viability, and lesions in lysosome membranes are repaired by the ESCRT machinery. Here, we describe an additional mechanism for lysosome repair that is activated independently of ESCRT recruitment. Lipidomic analyses showed increases in lysosomal phosphatidylserine and cholesterol after damage. Electron microscopy demonstrated that lysosomal membrane damage is rapidly followed by the formation of contacts with the endoplasmic reticulum (ER), which depends on the ER proteins VAPA/B. The cholesterol-binding protein ORP1L was recruited to damaged lysosomes, accompanied by cholesterol accumulation by a mechanism that required VAP–ORP1L interactions. The PtdIns 4-kinase PI4K2A rapidly produced PtdIns4P on lysosomes upon damage, and knockout of PI4K2A inhibited damage-induced accumulation of ORP1L and cholesterol and led to the failure of lysosomal membrane repair. The cholesterol–PtdIns4P transporter OSBP was also recruited upon damage, and its depletion caused lysosomal accumulation of PtdIns4P and resulted in cell death. We conclude that ER contacts are activated on damaged lysosomes in parallel to ESCRTs to provide lipids for membrane repair, and that PtdIns4P generation and removal are central in this response.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | e112677 |
| Tidsskrift | EMBO Journal |
| Vol/bind | 41 |
| Udgave nummer | 24 |
| ISSN | 0261-4189 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
The Core Facilities for Advanced Light Microscopy and Advanced Electron Microscopy at Oslo University Hospital are acknowledged for providing access to and training on relevant microscopes. We thank the Lipidomics Core Facility at the Danish Cancer Society Research Center for providing access to instrumentations and materials. We thank Ulrikke Dahl Brinch and Catherine Sem Wegner for excellent technical help with sample preparation for electron microscopy. We thank Pietro De Camilli for kindly providing VAPA/B knockout cells and Matthew Yoke Wui Ng and Kia Wee Tan for providing plasmid constructs. AHL was supported by a Young Research Talents Grant from the Research Council of Norway (project number 325305). CR was supported by a grant from the Norwegian Cancer Society (project number 198140). HS was supported by the Norwegian Cancer Society (project number 182698), the South‐Eastern Norway Regional Health Authority (project number 2016087), the Research Council of Norway (project number 302994), and the European Research Council (Advanced Grant number 788954). MJ was supported by grants from the Danish National Research Foundation (DNRF125) and the Novo Nordisk Foundation (NNF17OC0029432), and KM was supported by the Independent Research Fund Denmark (6108‐00542B). This work was partly supported by the Research Council of Norway through its Centres of Excellence funding scheme (project number 262652). Figures were created using Adobe Illustrator CS6 and BioRender ( https://biorender.com/ ).
Publisher Copyright:
© 2022 The Authors. Published under the terms of the CC BY NC ND 4.0 license.
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