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

Dmitry Zenko; Dawn Thompson; James N. Hislop

doi:10.1016/j.neuropharm.2019.107828

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

University of Oxford

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

14 Downloads (Pure)

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 language	English
Article number	107828
Number of pages	13
Journal	Neuropharmacology
Volume	162
Early online date	22 Oct 2019
DOIs	https://doi.org/10.1016/j.neuropharm.2019.107828
Publication status	Published - 1 Jan 2020

Bibliographical 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)

Keywords

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

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© 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
Accepted author manuscript, 4.69 MBLicence: CC BY-NC-ND

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

Abstract

Bibliographical note

Keywords

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James Hislop

Iain Fraser Cytometry Centre

Microscopy and Histology

Cite this

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

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

Profiles

James Hislop

Equipment

Iain Fraser Cytometry Centre

Microscopy and Histology

Cite this