Endocytic Sorting and Downregulation of the M2 Acetylcholine Receptor is Regulated by Ubiquitin and the ESCRT Complex

Dmitry Zenko, Dawn Thompson, James N. Hislop* (Corresponding Author)

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Cholinergic dysfunction plays a critical role in a number of disease states, and the loss of functional muscarinic acetylcholine receptors plays a key role in disease pathogenesis. Therefore, preventing receptor downregulation would maintain functional receptor number, and be predicted to alleviate symptoms. However, the molecular mechanism(s) underlying muscarinic receptor downregulation are currently unknown. Here we demonstrate that the M2 muscarinic receptor undergoes rapid lysosomal proteolysis, and this lysosomal trafficking is facilitated by ubiquitination of the receptor. Importantly, we show that this trafficking is driven specifically by ESCRT mediated involution. Critically, we provide evidence that disruption of this process leads to a re-routing of the trafficking of the M2 receptor away from the lysosome and into recycling pathway, and eventually back to the plasma membrane. This study is the first to identify the process by which the M2 muscarinic receptor undergoes endocytic sorting, and critically reveals a regulatory checkpoint that represents a target to pharmacologically increase the number of functional muscarinic receptors within the central nervous system.
Original languageEnglish
Article number107828
Number of pages13
JournalNeuropharmacology
Volume162
Early online date22 Oct 2019
DOIs
Publication statusPublished - 1 Jan 2020

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Endosomal Sorting Complexes Required for Transport
Cholinergic Receptors
Muscarinic Receptors
Ubiquitin
Muscarinic M2 Receptors
Down-Regulation
Ubiquitination
Lysosomes
Cholinergic Agents
Proteolysis
Central Nervous System
Cell Membrane

Keywords

  • Muscarinic Receptor
  • lysosome
  • ubiquitin
  • endosome
  • ESCRT
  • recycling
  • Ubiquitin
  • Lysosome
  • Muscarinic receptor
  • Recycling
  • Endosome

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Pharmacology

Cite this

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title = "Endocytic Sorting and Downregulation of the M2 Acetylcholine Receptor is Regulated by Ubiquitin and the ESCRT Complex",
abstract = "Cholinergic dysfunction plays a critical role in a number of disease states, and the loss of functional muscarinic acetylcholine receptors plays a key role in disease pathogenesis. Therefore, preventing receptor downregulation would maintain functional receptor number, and be predicted to alleviate symptoms. However, the molecular mechanism(s) underlying muscarinic receptor downregulation are currently unknown. Here we demonstrate that the M2 muscarinic receptor undergoes rapid lysosomal proteolysis, and this lysosomal trafficking is facilitated by ubiquitination of the receptor. Importantly, we show that this trafficking is driven specifically by ESCRT mediated involution. Critically, we provide evidence that disruption of this process leads to a re-routing of the trafficking of the M2 receptor away from the lysosome and into recycling pathway, and eventually back to the plasma membrane. This study is the first to identify the process by which the M2 muscarinic receptor undergoes endocytic sorting, and critically reveals a regulatory checkpoint that represents a target to pharmacologically increase the number of functional muscarinic receptors within the central nervous system.",
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note = "Acknowledgements The authors would like to thank Professor Mark von Zastrow from the University of California, San Francisco for sharing critical constructs. We would like to thank Kevin MacKenzie and the University of Aberdeen Microscopy core and the Iain Fraser Flow cytometry core for their assistance in the acquisition of data, and Professor Lynda Erskine for critical reading of the manuscript. This work was supported by a PhD studentship from the University of Aberdeen (DZ) and by funding from the Royal Society and Tenovus Scotland (JNH)",
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N1 - Acknowledgements The authors would like to thank Professor Mark von Zastrow from the University of California, San Francisco for sharing critical constructs. We would like to thank Kevin MacKenzie and the University of Aberdeen Microscopy core and the Iain Fraser Flow cytometry core for their assistance in the acquisition of data, and Professor Lynda Erskine for critical reading of the manuscript. This work was supported by a PhD studentship from the University of Aberdeen (DZ) and by funding from the Royal Society and Tenovus Scotland (JNH)

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N2 - Cholinergic dysfunction plays a critical role in a number of disease states, and the loss of functional muscarinic acetylcholine receptors plays a key role in disease pathogenesis. Therefore, preventing receptor downregulation would maintain functional receptor number, and be predicted to alleviate symptoms. However, the molecular mechanism(s) underlying muscarinic receptor downregulation are currently unknown. Here we demonstrate that the M2 muscarinic receptor undergoes rapid lysosomal proteolysis, and this lysosomal trafficking is facilitated by ubiquitination of the receptor. Importantly, we show that this trafficking is driven specifically by ESCRT mediated involution. Critically, we provide evidence that disruption of this process leads to a re-routing of the trafficking of the M2 receptor away from the lysosome and into recycling pathway, and eventually back to the plasma membrane. This study is the first to identify the process by which the M2 muscarinic receptor undergoes endocytic sorting, and critically reveals a regulatory checkpoint that represents a target to pharmacologically increase the number of functional muscarinic receptors within the central nervous system.

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