Abstract
While severe acute respiratory syndrome coronavirus (SARS-CoV) was initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that virus entry may also involve endocytosis. We have found that SARS-CoV enters cells via pH- and receptor-dependent endocytosis. Treatment of cells with either SARS-CoV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 (ACE2), the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic pathway inhibitors and dominant-negative Eps15 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research.
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Abbreviations
- (ACE2):
-
angiotensin converting enzyme 2
- (CPZ):
-
chlorpromazine
- (CTB):
-
cholera toxin subunit B
- (EEA1):
-
early endosome antigen 1
- (HEK293E):
-
human embryonic kidney 293E
- (MβCD):
-
methyl-β-cyclodextrin
- (SARS):
-
severe acute respiratory syndrome
- (SARS-CoV):
-
severe acute respiratory syndrome coronavirus
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Acknowledgements
We thank Dr James Kappler for editing. We thank Drs Brian Seed and Michael Farzan from Harvard Medical School, Dr Alexandre Benmerah (through Dr Du Feng, Tsinghua University, China) from Institut Pasteur for providing materials and reagents. We thank Ju Xiangwu from Shenyang Pharmaceutical University, China for the statistic analysis. This work was funded by National Natural Science Foundation of China (30421003, 30528002, 30625013, and 30623009) and Ministry of Science and Technology of China (2006AA02Z152 and 2005CB523000). Y. Z. was supported by NNSFC 30600020.
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Wang, H., Yang, P., Liu, K. et al. SARS coronavirus entry into host cells through a novel clathrin- and caveolae-independent endocytic pathway. Cell Res 18, 290–301 (2008). https://doi.org/10.1038/cr.2008.15
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DOI: https://doi.org/10.1038/cr.2008.15
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