Wuhan Institute of Virology Created COVID-19-Like Virus In 2007

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Difference in Receptor Usage between Severe Acute Respiratory Syndrome (SARS) Coronavirus and SARS-Like Coronavirus of Bat Origin
Wuze Ren, Xiuxia Qu, Wendong Li, Zhenggang Han, Meng Yu, Peng Zhou, Shu-Yi Zhang, Lin-Fa Wang, Hongkui Deng, Zhengli Shi
15 November 2007

ABSTRACT
Severe acute respiratory syndrome (SARS) is caused by the SARS-associated coronavirus (SARS-CoV), which uses angiotensin-converting enzyme 2 (ACE2) as its receptor for cell entry. A group of SARS-like CoVs (SL-CoVs) has been identified in horseshoe bats. SL-CoVs and SARS-CoVs share identical genome organizations and high sequence identities, with the main exception of the N terminus of the spike protein (S), known to be responsible for receptor binding in CoVs. In this study, we investigated the receptor usage of the SL-CoV S by combining a human immunodeficiency virus-based pseudovirus system with cell lines expressing the ACE2 molecules of human, civet, or horseshoe bat. In addition to full-length S of SL-CoV and SARS-CoV, a series of S chimeras was constructed by inserting different sequences of the SARS-CoV S into the SL-CoV S backbone. Several important observations were made from this study. First, the SL-CoV S was unable to use any of the three ACE2 molecules as its receptor. ...

... a minimal insert region (amino acids 310 to 518) was found to be sufficient to convert the SL-CoV S from non-ACE2 binding to human ACE2 binding
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In this study, a human immunodeficiency virus (HIV)-based pseudovirus system was employed to address these issues. Our results indicated that the SL-CoV S protein is unable to use ACE2 proteins of different species for cell entry and that SARS-CoV S protein also failed to bind the ACE2 molecule of the horseshoe bat, Rhinolophus pearsonii. However, when the RBD of SL-CoV S was replaced with that from the SARS-CoV S, the hybrid S protein was able to use the huACE2 for cell entry

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CREDITS
This work was jointly funded by a State Key Program for Basic Research Grant (2005CB523004) from the Chinese Ministry of Science and Technology, a special fund from the president of the Chinese Academy of Sciences (no. 1009), the Knowledge Innovation Program Key Project of the Chinese Academy of Sciences (KSCX1-YW-R-07) to Z. Shi, the Sixth Framework Program “EPISARS” of the European Commission, a National Nature Science Foundation of China for Creative Research group grant (30421004) to H. Deng, and the Australian Biosecurity CRC for Emerging Infectious Diseases (project 1.026RE) to L.-F. Wang.

Corresponding author. Mailing address for Z. Shi: State key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei 430071, China. Phone: (86-27)-87197240. Fax: (86-27)-87197240. E-mail: zlshi@wh.iov.cn.

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