Degron-tagged reporters probe membrane topology and enable the specific labelling of membrane-wrapped structures

Katharina B. Beer, Gholamreza Fazeli, Kristyna Judasova, Linda Irmisch, Jona Causemann, Jörg Mansfeld and Ann M. Wehman ()
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Katharina B. Beer: Julius-Maximilians-Universität Würzburg
Gholamreza Fazeli: Julius-Maximilians-Universität Würzburg
Kristyna Judasova: Dresden International Graduate School for Biomedicine and Bioengineering (DIGS-BB)
Linda Irmisch: Julius-Maximilians-Universität Würzburg
Jona Causemann: Julius-Maximilians-Universität Würzburg
Jörg Mansfeld: Technische Universität Dresden
Ann M. Wehman: Julius-Maximilians-Universität Würzburg

Nature Communications, 2019, vol. 10, issue 1, 1-16

Abstract: Abstract 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.

Date: 2019
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DOI: 10.1038/s41467-019-11442-z

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