tetano
Editor, Senior Moderator
Cell Chem Biol
. 2023 Feb 28;S2451-9456(23)00032-6.
doi: 10.1016/j.chembiol.2023.02.001. Online ahead of print.
SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
Lu Miao[SUP] 1 [/SUP], Chunyu Yan[SUP] 2 [/SUP], Yingzhu Chen[SUP] 1 [/SUP], Wei Zhou[SUP] 2 [/SUP], Xuelian Zhou[SUP] 2 [/SUP], Qinglong Qiao[SUP] 1 [/SUP], Zhaochao Xu[SUP] 3 [/SUP]
Affiliations
Abstract
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.
Keywords: COVID-19; Halo tag; RBD degradation; RBD-ACE2 interaction; SARS-CoV-2 infection; SIM imaging; SNAP tag; endocytosis; self-labeling; super-resolution imaging.
. 2023 Feb 28;S2451-9456(23)00032-6.
doi: 10.1016/j.chembiol.2023.02.001. Online ahead of print.
SIM imaging resolves endocytosis of SARS-CoV-2 spike RBD in living cells
Lu Miao[SUP] 1 [/SUP], Chunyu Yan[SUP] 2 [/SUP], Yingzhu Chen[SUP] 1 [/SUP], Wei Zhou[SUP] 2 [/SUP], Xuelian Zhou[SUP] 2 [/SUP], Qinglong Qiao[SUP] 1 [/SUP], Zhaochao Xu[SUP] 3 [/SUP]
Affiliations
- PMID: 36889309
- DOI: 10.1016/j.chembiol.2023.02.001
Abstract
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.
Keywords: COVID-19; Halo tag; RBD degradation; RBD-ACE2 interaction; SARS-CoV-2 infection; SIM imaging; SNAP tag; endocytosis; self-labeling; super-resolution imaging.