ER-export and ARFRP1/AP-1-dependent delivery of SARS-CoV-2 Envelope to lysosomes controls late stages of viral replication

Guy Pearson; Harriet Mears; Malgorzata Broncel; Ambrosius P. Snijders; David Bauer; Jeremy Carlton

doi:10.1126/sciadv.adl5012

ER-export and ARFRP1/AP-1-dependent delivery of SARS-CoV-2 Envelope to lysosomes controls late stages of viral replication

Guy Pearson, Harriet Mears, Malgorzata Broncel, Ambrosius P. Snijders, David Bauer, Jeremy Carlton^*

^*Corresponding author for this work

Comprehensive Cancer Centre

Francis Crick Institute

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

5 Citations (Scopus)

134 Downloads (Pure)

Abstract

The β-coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the global COVID-19 pandemic. Coronaviral Envelope (E) proteins are pentameric viroporins that play essential roles in assembly, release, and pathogenesis. We developed a nondisruptive tagging strategy for SARS-CoV-2 E and find that, at steady state, it localizes to the Golgi and to lysosomes. We identify sequences in E, conserved across Coronaviridae, responsible for endoplasmic reticulum–to–Golgi export, and relate this activity to interaction with COP-II via SEC24. Using proximity biotinylation, we identify an ADP ribosylation factor 1/adaptor protein–1 (ARFRP1/ AP-1)–dependent pathway allowing Golgi-to-lysosome trafficking of E. We identify sequences in E that bind AP-1, are conserved across β-coronaviruses, and allow E to be trafficked from Golgi to lysosomes. We show that E acts to deacidify lysosomes and, by developing a trans-complementation assay for SARS-CoV-2 structural proteins, that lysosomal delivery of E and its viroporin activity is necessary for efficient viral replication and release.

Original language	English
Article number	eadl5012
Journal	Science Advances
Volume	10
Issue number	14
DOIs	https://doi.org/10.1126/sciadv.adl5012
Publication status	Published - 3 Apr 2024

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1126/sciadv.adl5012

adl5012_Combined_Red_2
This research was funded in whole, or in part, by the Wellcome Trust (206346/Z/17/Z, 224484/Z/21/Z, CC1002, CC2166). For the purpose of Open Access, the authors have applied a Creative Commons Attribution (CC BY) public copyright licence to any Author Accepted Manuscript version arising from this submission.
Accepted author manuscript, 18.8 MBLicence: CC BY

Cite this

@article{6ac8f82631e74bf78ad7ed0bf759a4a0,

title = "ER-export and ARFRP1/AP-1-dependent delivery of SARS-CoV-2 Envelope to lysosomes controls late stages of viral replication",

abstract = "The β-coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the global COVID-19 pandemic. Coronaviral Envelope (E) proteins are pentameric viroporins that play essential roles in assembly, release, and pathogenesis. We developed a nondisruptive tagging strategy for SARS-CoV-2 E and find that, at steady state, it localizes to the Golgi and to lysosomes. We identify sequences in E, conserved across Coronaviridae, responsible for endoplasmic reticulum–to–Golgi export, and relate this activity to interaction with COP-II via SEC24. Using proximity biotinylation, we identify an ADP ribosylation factor 1/adaptor protein–1 (ARFRP1/ AP-1)–dependent pathway allowing Golgi-to-lysosome trafficking of E. We identify sequences in E that bind AP-1, are conserved across β-coronaviruses, and allow E to be trafficked from Golgi to lysosomes. We show that E acts to deacidify lysosomes and, by developing a trans-complementation assay for SARS-CoV-2 structural proteins, that lysosomal delivery of E and its viroporin activity is necessary for efficient viral replication and release.",

author = "Guy Pearson and Harriet Mears and Malgorzata Broncel and Snijders, {Ambrosius P.} and David Bauer and Jeremy Carlton",

note = "Article is accepted in principle and under technical hold to allow APC payment. I've put a month-long embargo on the files, but will remove this and release it when we get final acceptance as this is OA through a CCBY license with the relevant RRS applied",

year = "2024",

month = apr,

day = "3",

doi = "10.1126/sciadv.adl5012",

language = "English",

volume = "10",

journal = "Science Advances",

issn = "2375-2548",

publisher = "American Association for the Advancement of Science",

number = "14",

}

TY - JOUR

T1 - ER-export and ARFRP1/AP-1-dependent delivery of SARS-CoV-2 Envelope to lysosomes controls late stages of viral replication

AU - Pearson, Guy

AU - Mears, Harriet

AU - Broncel, Malgorzata

AU - Snijders, Ambrosius P.

AU - Bauer, David

AU - Carlton, Jeremy

N1 - Article is accepted in principle and under technical hold to allow APC payment. I've put a month-long embargo on the files, but will remove this and release it when we get final acceptance as this is OA through a CCBY license with the relevant RRS applied

PY - 2024/4/3

Y1 - 2024/4/3

N2 - The β-coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the global COVID-19 pandemic. Coronaviral Envelope (E) proteins are pentameric viroporins that play essential roles in assembly, release, and pathogenesis. We developed a nondisruptive tagging strategy for SARS-CoV-2 E and find that, at steady state, it localizes to the Golgi and to lysosomes. We identify sequences in E, conserved across Coronaviridae, responsible for endoplasmic reticulum–to–Golgi export, and relate this activity to interaction with COP-II via SEC24. Using proximity biotinylation, we identify an ADP ribosylation factor 1/adaptor protein–1 (ARFRP1/ AP-1)–dependent pathway allowing Golgi-to-lysosome trafficking of E. We identify sequences in E that bind AP-1, are conserved across β-coronaviruses, and allow E to be trafficked from Golgi to lysosomes. We show that E acts to deacidify lysosomes and, by developing a trans-complementation assay for SARS-CoV-2 structural proteins, that lysosomal delivery of E and its viroporin activity is necessary for efficient viral replication and release.

AB - The β-coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the global COVID-19 pandemic. Coronaviral Envelope (E) proteins are pentameric viroporins that play essential roles in assembly, release, and pathogenesis. We developed a nondisruptive tagging strategy for SARS-CoV-2 E and find that, at steady state, it localizes to the Golgi and to lysosomes. We identify sequences in E, conserved across Coronaviridae, responsible for endoplasmic reticulum–to–Golgi export, and relate this activity to interaction with COP-II via SEC24. Using proximity biotinylation, we identify an ADP ribosylation factor 1/adaptor protein–1 (ARFRP1/ AP-1)–dependent pathway allowing Golgi-to-lysosome trafficking of E. We identify sequences in E that bind AP-1, are conserved across β-coronaviruses, and allow E to be trafficked from Golgi to lysosomes. We show that E acts to deacidify lysosomes and, by developing a trans-complementation assay for SARS-CoV-2 structural proteins, that lysosomal delivery of E and its viroporin activity is necessary for efficient viral replication and release.

UR - http://www.scopus.com/inward/record.url?scp=85190076906&partnerID=8YFLogxK

U2 - 10.1126/sciadv.adl5012

DO - 10.1126/sciadv.adl5012

M3 - Article

SN - 2375-2548

VL - 10

JO - Science Advances

JF - Science Advances

IS - 14

M1 - eadl5012

ER -

ER-export and ARFRP1/AP-1-dependent delivery of SARS-CoV-2 Envelope to lysosomes controls late stages of viral replication

Abstract

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this