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SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells

It is urgent to understand the infection mechanism of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) for the prevention and treatment of COVID-19. The infection of SARS-CoV-2 starts when the receptor-binding domain (RBD) of viral spike protein binds to angiotensin-converting enzyme 2 (...

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Autores principales: Miao, Lu, Yan, Chunyu, Chen, Yingzhu, Zhou, Wei, Zhou, Xuelian, Qiao, Qinglong, Xu, Zhaochao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Ltd. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9990177/
https://www.ncbi.nlm.nih.gov/pubmed/36889309
http://dx.doi.org/10.1016/j.chembiol.2023.02.001
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author Miao, Lu
Yan, Chunyu
Chen, Yingzhu
Zhou, Wei
Zhou, Xuelian
Qiao, Qinglong
Xu, Zhaochao
author_facet Miao, Lu
Yan, Chunyu
Chen, Yingzhu
Zhou, Wei
Zhou, Xuelian
Qiao, Qinglong
Xu, Zhaochao
author_sort Miao, Lu
collection PubMed
description It is urgent to understand the infection mechanism of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) for the prevention and treatment of COVID-19. The infection of SARS-CoV-2 starts when the receptor-binding domain (RBD) of viral spike protein binds to angiotensin-converting enzyme 2 (ACE2) of the host cell, but the endocytosis details after this binding are not clear. Here, RBD and ACE2 were genetically coded and labeled with organic dyes to track RBD endocytosis in living cells. The photostable dyes enable long-term structured illumination microscopy (SIM) imaging and to quantify RBD-ACE2 binding (RAB) by the intensity ratio of RBD/ACE2 fluorescence. We resolved RAB endocytosis in living cells, including RBD-ACE2 recognition, cofactor-regulated membrane internalization, RAB-bearing vesicle formation and transport, RAB degradation, and downregulation of ACE2. The RAB was found to activate the RBD internalization. After vesicles were transported and matured within cells, RAB was finally degraded after being taken up by lysosomes. This strategy is a promising tool to understand the infection mechanism of SARS-CoV-2.
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spelling pubmed-99901772023-03-08 SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells Miao, Lu Yan, Chunyu Chen, Yingzhu Zhou, Wei Zhou, Xuelian Qiao, Qinglong Xu, Zhaochao Cell Chem Biol Article It is urgent to understand the infection mechanism of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) for the prevention and treatment of COVID-19. The infection of SARS-CoV-2 starts when the receptor-binding domain (RBD) of viral spike protein binds to angiotensin-converting enzyme 2 (ACE2) of the host cell, but the endocytosis details after this binding are not clear. Here, RBD and ACE2 were genetically coded and labeled with organic dyes to track RBD endocytosis in living cells. The photostable dyes enable long-term structured illumination microscopy (SIM) imaging and to quantify RBD-ACE2 binding (RAB) by the intensity ratio of RBD/ACE2 fluorescence. We resolved RAB endocytosis in living cells, including RBD-ACE2 recognition, cofactor-regulated membrane internalization, RAB-bearing vesicle formation and transport, RAB degradation, and downregulation of ACE2. The RAB was found to activate the RBD internalization. After vesicles were transported and matured within cells, RAB was finally degraded after being taken up by lysosomes. This strategy is a promising tool to understand the infection mechanism of SARS-CoV-2. Elsevier Ltd. 2023-03-16 2023-03-07 /pmc/articles/PMC9990177/ /pubmed/36889309 http://dx.doi.org/10.1016/j.chembiol.2023.02.001 Text en © 2023 Elsevier Ltd. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Miao, Lu
Yan, Chunyu
Chen, Yingzhu
Zhou, Wei
Zhou, Xuelian
Qiao, Qinglong
Xu, Zhaochao
SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
title SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
title_full SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
title_fullStr SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
title_full_unstemmed SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
title_short SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
title_sort sim imaging resolves endocytosis of sars-cov-2 spike rbd in living cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9990177/
https://www.ncbi.nlm.nih.gov/pubmed/36889309
http://dx.doi.org/10.1016/j.chembiol.2023.02.001
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