【摘 要】
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Bat coronavirus (CoV) RaTG13 shares the highest genome sequence identity with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among all known coronaviruses,and also uses human angiotensin converting enzyme 2 (hACE2) for virus entry.Thus,SARS-
【机 构】
：
NHC Key Laboratory of Systems Biology of Pathogens,Institute of Pathogen Biology,Chinese Academy of
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Bat coronavirus (CoV) RaTG13 shares the highest genome sequence identity with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among all known coronaviruses,and also uses human angiotensin converting enzyme 2 (hACE2) for virus entry.Thus,SARS-CoV-2 is thought to have originated from bat.However,whether SARS-CoV-2 emerged from bats directly or through an intermediate host remains elusive.Here,we found that Rhinolophus affinis bat ACE2 (RaACE2) is an entry receptor for both SARS-CoY-2 and RaTG13,although the binding of RaACE2 to the receptor-binding domain (RBD) of SARS-CoV-2 is markedly weaker than that of hACE2.We further evaluated the receptor activities of ACE2s from additional 16 diverse animal species for RaTG13,SARS-CoV,and SARS-CoV-2 in terms of S protein bind-ing,membrane fusion,and pseudovirus entry.We found that the RaTG 13 spike (S) protein is significantly less fusogenic than SARS-CoV and SARS-CoV-2,and seven out of sixteen different ACE2s function as entry receptors for all three viruses,indicating that all three viruses might have broad host rages.Of note,RaTG13 S pseudovirions can use mouse,but not pangolin ACE2,for virus entry,whereas SARS-CoV-2 S pseudovirions can use pangolin,but not mouse,ACE2 enter cells efficiently.Mutagenesis analysis revealed that residues 484 and 498 in RaTG13 and SARS-CoV-2 S proteins play critical roles in recognition of mouse and human ACE2s.Finally,two polymorphous Rhinolophous sinicus bat ACE2s showed different susceptibilities to virus entry by RaTG13 and SARS-CoV-2 S pseudovirions,suggesting possible coevolu-tion.Our results offer better understanding of the mechanism of coronavirus entry,host range,and virus-host coevolution.
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