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

Nature Communications
Katharina B BeerAnn M Wehman

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.

References

May 1, 1974·Genetics·S Brenner
Jan 1, 1996·Annual Review of Cell and Developmental Biology·R D Teasdale, M R Jackson
May 3, 2003·Nature Reviews. Molecular Cell Biology·Michael Edidin
Jun 28, 2003·Methods : a Companion to Methods in Enzymology·Ravi S Kamath, Julie Ahringer
Feb 21, 2006·Nature Methods·Holger LorenzJennifer Lippincott-Schwartz
May 3, 2007·Pflügers Archiv : European journal of physiology·Péter Várnai, Tamas Balla
Oct 25, 2007·Journal of Proteome Research·Dong-Sic ChoiYong Song Gho
Jan 1, 2009·Nature Methods·Stefan W Hell
Oct 21, 2009·Nature Chemical Biology·Erin K SchraderAndreas Matouschek
Nov 17, 2009·Nature Methods·Kohei NishimuraMasato Kanemaki
Feb 1, 2011·Nature Methods·Stefanie RedemannHenrik Bringmann
Sep 14, 2011·Annual Review of Pathology·Ronald S FlannaganSergio Grinstein
Nov 22, 2011·Current Biology : CB·Ann M WehmanJeremy Nance
Dec 14, 2011·Nature Structural & Molecular Biology·Emmanuel CaussinusMarkus Affolter
Jan 22, 2013·Trends in Neurosciences·Tomoko YamadaAzad Bonni
Nov 13, 2013·The Journal of Cell Biology·Rebecca A GreenKaren Oegema
Sep 17, 2015·Scientific Reports·Yeong Ju ShinByung Joon Hwang
Feb 10, 2016·Proceedings of the National Academy of Sciences of the United States of America·Joanna KowalClotilde Théry
Mar 26, 2016·Current Opinion in Cell Biology·Ramon D Jones, Richard G Gardner
Aug 26, 2016·Immunological Reviews·Roni LevinJohnathan Canton
Oct 1, 2016·Cell Adhesion & Migration·Katharina B Beer, Ann Marie Wehman
Dec 10, 2016·Physiological Reviews·Natalie FootSharad Kumar
Jan 26, 2017·Nature Reviews. Molecular Cell Biology·Rosemarie Ungricht, Ulrike Kutay
Mar 23, 2017·The Journal of Cell Biology·Julia KönigThomas Müller-Reichert
Nov 28, 2017·Annual Review of Genetics·Toyoaki Natsume, Masato T Kanemaki
Jan 8, 2018·Current Opinion in Cell Biology·Larry Gerace, Olga Tapia
Jan 9, 2018·Trends in Cell Biology·Henri-François RenardPierre Morsomme
Jan 18, 2018·Nature Reviews. Molecular Cell Biology·Guillaume van NielGraça Raposo
Jan 26, 2018·Proceedings of the National Academy of Sciences of the United States of America·Katharina B BeerAnn M Wehman

❮ Previous
Next ❯

Citations

Mar 19, 2020·Angewandte Chemie·Fangfei YinYing Zhu
Aug 15, 2020·Frontiers in Cell and Developmental Biology·Gholamreza FazeliAnn M Wehman
Jul 1, 2021·Proceedings of the National Academy of Sciences of the United States of America·Xinya HuangXuezhu Feng
Aug 28, 2021·Nature Methods·Frederik J VerweijGuillaume van Niel

❮ Previous
Next ❯

Methods Mentioned

BETA
light microscopy
ubiquitination
transgenic
electron tomography
fluorescence recovery after photobleaching
light
electron microscopy
Transfection
PCR
PCRs

Software Mentioned

Adobe Photoshop
Excel
ImageJ
OriginPro
Fiji
Leica LAS X
OriginLab
Photoshop
Prism
Leica LAS

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Hospital Trustee
J Schenker
Methods : a Companion to Methods in Enzymology
Hongning WangAlfred L Fisher
Current Protocols in Chemical Biology
Maya T Kunkel, Alexandra C Newton
© 2021 Meta ULC. All rights reserved