Degron-tagged reporters probe membrane topology and enable the specific labelling of membrane-wrapped structures.
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Date
2019-08-02ICR Author
Author
Beer, KB
Fazeli, G
Judasova, K
Irmisch, L
Causemann, J
Mansfeld, J
Wehman, AM
Type
Journal Article
Metadata
Show full item recordAbstract
Visualization of specific organelles in tissues over background fluorescence can be challenging, especially when reporters localize to multiple structures. Instead of trying to identify proteins enriched in specific membrane-wrapped structures, we use a selective degradation approach to remove reporters from the cytoplasm or nucleus of C. elegans embryos and mammalian cells. We demonstrate specific labelling of organelles using degron-tagged reporters, including extracellular vesicles, as well as individual neighbouring membranes. These degron-tagged reporters facilitate long-term tracking of released cell debris and cell corpses, even during uptake and phagolysosomal degradation. We further show that degron protection assays can probe the topology of the nuclear envelope and plasma membrane during cell division, giving insight into protein and organelle dynamics. As endogenous and heterologous degrons are used in bacteria, yeast, plants, and animals, degron approaches can enable the specific labelling and tracking of proteins, vesicles, organelles, cell fragments, and cells in many model systems.
Collections
Subject
Hela Cells
Cell Membrane
Embryo, Nonmammalian
Animals
Humans
Caenorhabditis elegans
Luminescent Proteins
Staining and Labeling
Genes, Reporter
Fluorescence
Proteolysis
Extracellular Vesicles
Intravital Microscopy
Research team
Post-translational modifications and cell proliferation
Language
eng
Date accepted
2019-07-16
License start date
2019-08-02
Citation
Nature communications, 2019, 10 (1), pp. 3490 - ?
Publisher
NATURE PUBLISHING GROUP