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Characterization of the 3a protein of SARS-associated coronavirus in infected vero E6 cells and SARS patients.

ABSTRACT: Proteomics was used to identify a protein encoded by ORF 3a in a SARS-associated coronavirus (SARS-CoV). Immuno-blotting revealed that interchain disulfide bonds might be formed between this protein and the spike protein. ELISA indicated that sera from SARS patients have significant positive reactions with synthesized peptides derived from the 3a protein. These results are concordant with that of a spike protein-derived peptide. A tendency exists for co-mutation between the 3a protein and the spike protein of SARS-CoV isolates, suggesting that the function of the 3a protein correlates with the spike protein. Taken together, the 3a protein might be tightly correlated to the spike protein in the SARS-CoV functions. The 3a protein may serve as a new clinical marker or drug target for SARS treatment.

SUBMITTER: Zeng R

PROVIDER: S-EPMC7127270 | biostudies-literature | 2004 Jul

REPOSITORIES: biostudies-literature

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Characterization of the 3a protein of SARS-associated coronavirus in infected vero E6 cells and SARS patients.

Zeng Rong R Yang Rui-Fu RF Shi Mu-De MD Jiang Man-Rong MR Xie You-Hua YH Ruan Hong-Qiang HQ Jiang Xiao-Sheng XS Shi Lv L Zhou Hu H Zhang Lei L Wu Xiao-Dong XD Lin Ying Y Ji Yong-Yong YY Xiong Lei L Jin Yan Y Dai Er-Hei EH Wang Xiao-Yi XY Si Bin-Ying BY Wang Jin J Wang Hong-Xia HX Wang Cui-E CE Gan Yong-Hua YH Li Yu-Chuan YC Cao Ju-Tian JT Zuo Jiang-Ping JP Shan Shi-Fang SF Xie En E Chen Song-Hua SH Jiang Zhi-Qin ZQ Zhang Xi X Wang Yuan Y Pei Gang G Sun Bing B Wu Jia-Rui JR

Journal of molecular biology 20040701 1

Proteomics was used to identify a protein encoded by ORF 3a in a SARS-associated coronavirus (SARS-CoV). Immuno-blotting revealed that interchain disulfide bonds might be formed between this protein and the spike protein. ELISA indicated that sera from SARS patients have significant positive reactions with synthesized peptides derived from the 3a protein. These results are concordant with that of a spike protein-derived peptide. A tendency exists for co-mutation between the 3a protein and the sp ...[more]

PMID: 15312778

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Project description:BackgroundThe coronaviruses (CoVs) called the attention of the world for causing outbreaks of severe acute respiratory syndrome (SARS-CoV), in Asia in 2002-03, and respiratory disease in the Middle East (MERS-CoV), in 2012. In December 2019, yet again a new coronavirus (SARS-CoV-2) first identified in Wuhan, China, was associated with a severe respiratory infection, known today as COVID-19. This new virus quickly spread throughout China and 30 additional countries. As result, the World Health Organization (WHO) elevated the status of the COVID-19 outbreak from emergency of international concern to pandemic on March 11, 2020. The impact of COVID-19 on public health and economy fueled a worldwide race to approve therapeutic and prophylactic agents, but so far, there are no specific antiviral drugs or vaccines available. In current scenario, the development of in vitro systems for viral mass production and for testing antiviral and vaccine candidates proves to be an urgent matter.ObjectiveThe objective of this paper is study the biology of SARS-CoV-2 in Vero-E6 cells at the ultrastructural level.MethodsIn this study, we documented, by transmission electron microscopy and real-time reverse transcription polymerase chain reaction (RT-PCR), the infection of Vero-E6 cells with SARS-CoV-2 samples isolated from Brazilian patients.FindingsThe infected cells presented cytopathic effects and SARS-CoV-2 particles were observed attached to the cell surface and inside cytoplasmic vesicles. The entry of the virus into cells occurred through the endocytic pathway or by fusion of the viral envelope with the cell membrane. Assembled nucleocapsids were verified inside rough endoplasmic reticulum cisterns (RER). Viral maturation seemed to occur by budding of viral particles from the RER into smooth membrane vesicles.Main conclusionsTherefore, the susceptibility of Vero-E6 cells to SARS-CoV-2 infection and the viral pathway inside the cells were demonstrated by ultrastructural analysis.

Dataset Information

Characterization of the 3a protein of SARS-associated coronavirus in infected vero E6 cells and SARS patients.

Publications

Characterization of the 3a protein of SARS-associated coronavirus in infected vero E6 cells and SARS patients.

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