Loss of furin site enhances SARS-CoV-2 spike protein pseudovirus infection

Wang, Zeng; Zhong, Kunhong; Wang, Guoqing; Lu, Qizhong; Li, Hexian; Wu, Zhiguo; Zhang, Zongliang; Yang, Nian; Zheng, Meijun; Wang, Yuelong; Nie, Chao; Zhou, Liangxue; Tong, Aiping

doi:10.1016/j.gene.2022.147144

Cited by 7 publications

(5 citation statements)

References 42 publications

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“…The presence of the RBD on the C-terminal also allows the spike proteins to change conformations, promoting dissociation between the two subunits. Furthermore, SARS-CoV-2 spike proteins are unique because they contain an additional cleavage site, which has been shown to enhance infection [36]. The specific mechanisms explaining why MHV or SARS-CoV-1 entry into cells via macropinocytosis is limited are currently unknown, but the above-described structural and functional differences in spike proteins may explain differences in infection rate and host cell entry.…”

Section: Discussionmentioning

confidence: 99%

SARS-CoV-2 Spike Protein Stimulates Macropinocytosis in Murine and Human Macrophages via PKC-NADPH Oxidase Signaling

Ahn,

Burnett,

Pandey

et al. 2024

Antioxidants

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Coronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). While recent studies have demonstrated that SARS-CoV-2 may enter kidney and colon epithelial cells by inducing receptor-independent macropinocytosis, it remains unknown whether this process also occurs in cell types directly relevant to SARS-CoV-2-associated lung pneumonia, such as alveolar epithelial cells and macrophages. The goal of our study was to investigate the ability of SARS-CoV-2 spike protein subunits to stimulate macropinocytosis in human alveolar epithelial cells and primary human and murine macrophages. Flow cytometry analysis of fluid-phase marker internalization demonstrated that SARS-CoV-2 spike protein subunits S1, the receptor-binding domain (RBD) of S1, and S2 stimulate macropinocytosis in both human and murine macrophages in an angiotensin-converting enzyme 2 (ACE2)-independent manner. Pharmacological and genetic inhibition of macropinocytosis substantially decreased spike-protein-induced fluid-phase marker internalization in macrophages both in vitro and in vivo. High-resolution scanning electron microscopy (SEM) imaging confirmed that spike protein subunits promote the formation of membrane ruffles on the dorsal surface of macrophages. Mechanistic studies demonstrated that SARS-CoV-2 spike protein stimulated macropinocytosis via NADPH oxidase 2 (Nox2)-derived reactive oxygen species (ROS) generation. In addition, inhibition of protein kinase C (PKC) and phosphoinositide 3-kinase (PI3K) in macrophages blocked SARS-CoV-2 spike-protein-induced macropinocytosis. To our knowledge, these results demonstrate for the first time that SARS-CoV-2 spike protein subunits stimulate macropinocytosis in macrophages. These results may contribute to a better understanding of SARS-CoV-2 infection and COVID-19 pathogenesis.

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Section: Discussionmentioning

confidence: 99%

SARS-CoV-2 Spike Protein Stimulates Macropinocytosis in Murine and Human Macrophages via PKC-NADPH Oxidase Signaling

Ahn,

Burnett,

Pandey

et al. 2024

Antioxidants

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“…This oligopeptide is located next to the S1/S2 cleavage site [ 40 , 41 , 42 ]. In SARS-CoV-2 infected patients, the S-protein is cleaved into S1 and S2 [ 43 , 44 , 45 , 46 ], which is necessary for the virus’s penetration into the target cells. We suggest that the recognition of this site followed by hydrolysis will block S1/S2 proteolysis and neutralize the whole virus.…”

Section: Discussionmentioning

confidence: 99%

Natural Antibodies Produced in Vaccinated Patients and COVID-19 Convalescents Recognize and Hydrolyze Oligopeptides Corresponding to the S-Protein of SARS-CoV-2

Timofeeva,

Sedykh,

Sedykh

et al. 2023

Vaccines

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The S-protein is the major antigen of the SARS-CoV-2 virus, against which protective antibodies are generated. The S-protein gene was used in adenoviral vectors and mRNA vaccines against COVID-19. While the primary function of antibodies is to bind to antigens, catalytic antibodies can hydrolyze various substrates, including nucleic acids, proteins, oligopeptides, polysaccharides, and some other molecules. In this study, antibody fractions with affinity for RBD and S-protein (RBD-IgG and S-IgG) were isolated from the blood of COVID-19 patients vaccinated with Sputnik V. The fractions were analyzed for their potential to hydrolyze 18-mer oligopeptides corresponding to linear fragments of the SARS-CoV-2 S-protein. Here, we show that the IgG antibodies hydrolyze six out of nine oligopeptides efficiently, with the antibodies of COVID-19-exposed donors demonstrating the most significant activity. The IgGs of control donors not exposed to SARS-CoV-2 were found to be inactive in oligopeptide hydrolysis. The antibodies of convalescents and vaccinated patients were found to hydrolyze oligopeptides in a wide pH range, with the optimal pH range between 6.5 and 7.5. The hydrolysis of most oligopeptides by RBD-IgG antibodies is inhibited by thiol protease inhibitors, whereas S-IgG active centers generally combine several types of proteolytic activities. Ca2+ ions increase the catalytic activity of IgG preparations containing metalloprotease-like active centers. Thus, the proteolytic activity of natural antibodies against the SARS-CoV-2 protein is believed to be due to the similarity of catalytic antibodies’ active centers to canonical proteases. This work raises the question of the possible physiological role of proteolytic natural RBD-IgG and S-IgG resulting from vaccination and exposure to COVID-19.

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“… Johnson et al (2020) found that deletion of the last 19 amino acids of the cytoplasmic tail of the S protein, D614G and R682Q mutations all enhanced SARS-CoV-2 pseudotyped HIV-1 particle production and did not affect the neutralization ability against serum-neutralizing antibodies. Wang et al (2023b) also reported that deletion of the furan site on the S protein gene increased the pseudovirus’s yield and infection efficiency. SARS-CoV-2 pseudotyped VSV particles with G614 S protein mutation have higher titer than the corresponding D614 pseudovirus ( Korber et al, 2020 ).…”

Section: Survey Methodologymentioning

confidence: 96%

The development and application of pseudoviruses: assessment of SARS-CoV-2 pseudoviruses

Tan,

Wang,

Deng

et al. 2023

PeerJ

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Although most Coronavirus disease (COVID-19) patients can recover fully, the disease remains a significant cause of morbidity and mortality. In addition to the consequences of acute infection, a proportion of the population experiences long-term adverse effects associated with SARS-CoV-2. Therefore, it is still critical to comprehend the virus’s characteristics and how it interacts with its host to develop effective drugs and vaccines against COVID-19. SARS-CoV-2 pseudovirus, a replication-deficient recombinant glycoprotein chimeric viral particle, enables investigations of highly pathogenic viruses to be conducted without the constraint of high-level biosafety facilities, considerably advancing virology and being extensively employed in the study of SARS-CoV-2. This review summarizes three methods of establishing SARS-CoV-2 pseudovirus and current knowledge in vaccine development, neutralizing antibody research, and antiviral drug screening, as well as recent progress in virus entry mechanism and susceptible cell screening. We also discuss the potential advantages and disadvantages.

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Loss of furin site enhances SARS-CoV-2 spike protein pseudovirus infection

Cited by 7 publications

References 42 publications

SARS-CoV-2 Spike Protein Stimulates Macropinocytosis in Murine and Human Macrophages via PKC-NADPH Oxidase Signaling

SARS-CoV-2 Spike Protein Stimulates Macropinocytosis in Murine and Human Macrophages via PKC-NADPH Oxidase Signaling

Natural Antibodies Produced in Vaccinated Patients and COVID-19 Convalescents Recognize and Hydrolyze Oligopeptides Corresponding to the S-Protein of SARS-CoV-2

The development and application of pseudoviruses: assessment of SARS-CoV-2 pseudoviruses

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