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Journal of Virology, December 2008, p. 11985-11991, Vol. 82, No. 23
0022-538X/08/$08.00+0     doi:10.1128/JVI.01412-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Entry from the Cell Surface of Severe Acute Respiratory Syndrome Coronavirus with Cleaved S Protein as Revealed by Pseudotype Virus Bearing Cleaved S Protein{triangledown}

Rie Watanabe,1 Shutoku Matsuyama,1 Kazuya Shirato,1 Masami Maejima,1 Shuetsu Fukushi,2 Shigeru Morikawa,2 and Fumihiro Taguchi1*

Departments of Virology III,1 I, National Institute of Infectious Diseases, Murayama Branch, Musashi-Murayama, Tokyo 208-0011, Japan2

Received 8 July 2008/ Accepted 3 September 2008

Severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) is known to take an endosomal pathway for cell entry; however, it is thought to enter directly from the cell surface when a receptor-bound virion spike (S) protein is affected by trypsin, which induces cleavage of the S protein and activates its fusion potential. This suggests that SARS-CoV bearing a cleaved form of the S protein can enter cells directly from the cell surface without trypsin treatment. To explore this possibility, we introduced a furin-like cleavage sequence in the S protein at amino acids 798 to 801 and found that the mutated S protein was cleaved and induced cell fusion without trypsin treatment when expressed on the cell surface. Furthermore, a pseudotype virus bearing a cleaved S protein was revealed to infect cells in the presence of a lysosomotropic agent as well as a protease inhibitor, both of which are known to block SARS-CoV infection via an endosome, whereas the infection of pseudotypes with an uncleaved, wild-type S protein was blocked by these agents. A heptad repeat peptide, derived from a SARS-CoV S protein that is known to efficiently block infections from the cell surface, blocked the infection by a pseudotype with a cleaved S protein but not that with an uncleaved S protein. Those results indicate that SARS-CoV with a cleaved S protein is able to enter cells directly from the cell surface and agree with the previous observation of the protease-mediated cell surface entry of SARS-CoV.

* Corresponding author. Mailing address: Laboratory of Respiratory Viral Infections and SARS, National Institute of Infectious Diseases, Murayama Branch, 4-7-1 Gakuen, Mushai-Murayama, Tokyo 208-0011, Japan. Phone: 81-42-561-0771, ext. 3533. Fax: 81-42-567-5631. E-mail: ftaguchi{at}nih.go.jp

{triangledown} Published ahead of print on 10 September 2008.

Journal of Virology, December 2008, p. 11985-11991, Vol. 82, No. 23
0022-538X/08/$08.00+0     doi:10.1128/JVI.01412-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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