Document Detail


SARS-CoV nucleocapsid protein induced apoptosis of COS-1 mediated by the mitochondrial pathway.
MedLine Citation:
PMID:  17453707     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To investigate the apoptosis effect of SARS coronavirus nucleocapsid protein on cultured cell lines and to explore the possible pathway of apoptosis. pCDNA3.1(-)/his-myc vector containing the SARS coronavirus nucleocapsid gene (N), matric gene (M), spike gene (S) were transfected into COS-1, Huh-7 and HepG2 cells. Apoptosis induced by SARS coronavirus N protein under starvation of serum of COS-1 cells was monitored by Annexin V and electron microscopy assays. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (DeltaPsim) were determined by flow cytometric assay. Cytochrome C, cleaved caspase (cysteine aspartic acid protease)-3, 9, and poly (ADP-ribose) polymerase (PARP) were detected by Western blot. After removal of serum in COS-1 cells, we observed the loss of DeltaPsim, the increase of ROS and cytochrome C release into cytosol and subsequent activation of caspase-3 and PARP cleavage. The pan-caspase inhibitor z-VAD-fmk can block the activation of caspase 3, 9 and PARP cleavage. In conclusion, SARS coronavirus N protein can induce apoptosis of COS-1 cells by activating mitochondrial pathway. SARS coronavirus M, S protein can not induce apoptosis in COS-1, HepG2 and Huh-7 and SARS coronavirus N protein can not induce apoptosis in HepG2 and Huh-7 by methods used in this study.
Authors:
Lu Zhang; Lai Wei; Dong Jiang; Jianghua Wang; Xu Cong; Ran Fei
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Artificial cells, blood substitutes, and immobilization biotechnology     Volume:  35     ISSN:  1073-1199     ISO Abbreviation:  Artif Cells Blood Substit Immobil Biotechnol     Publication Date:  2007  
Date Detail:
Created Date:  2007-04-24     Completed Date:  2007-05-23     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9431307     Medline TA:  Artif Cells Blood Substit Immobil Biotechnol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  237-53     Citation Subset:  IM    
Affiliation:
Hepatology Institute, Peking University, People's Hospital, Beijing, People's Republic of China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis*
Blotting, Western
COS Cells
Carcinoma, Hepatocellular
Caspase 9 / metabolism
Cell Line, Tumor
Cercopithecus aethiops
Cytochromes c / metabolism
Flow Cytometry
Genetic Vectors
Humans
Liver Neoplasms
Membrane Glycoproteins / genetics,  metabolism
Membrane Potential, Mitochondrial / physiology
Microscopy, Electron
Mitochondria / physiology*,  ultrastructure,  virology*
Nucleocapsid Proteins / genetics*,  metabolism
Phosphatidylserines / metabolism
Reactive Oxygen Species / metabolism
SARS Virus / genetics*
Transfection
Viral Envelope Proteins / genetics,  metabolism
Chemical
Reg. No./Substance:
0/Membrane Glycoproteins; 0/Nucleocapsid Proteins; 0/Phosphatidylserines; 0/Reactive Oxygen Species; 0/Viral Envelope Proteins; 0/nucleocapsid protein, Coronavirus; 107476-75-5/spike glycoprotein, coronavirus; 9007-43-6/Cytochromes c; EC 3.4.22.-/Caspase 9

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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