ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat Cells
ABSTRACT Hundreds of sarbecoviruses have been found in bats, but only a fraction of them have the ability to infect cells using angiotensin-converting enzyme 2 (ACE2), the receptor for SARS-CoV and -2. To date, only ACE2-dependent sarbecoviruses have been isolated from field samples or grown in the...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2022-12-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.02566-22 |
| _version_ | 1811196275906838528 |
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| author | Hua Guo Ang Li Tian-Yi Dong Jia Su Yu-Lin Yao Yan Zhu Zheng-Li Shi Michael Letko |
| author_facet | Hua Guo Ang Li Tian-Yi Dong Jia Su Yu-Lin Yao Yan Zhu Zheng-Li Shi Michael Letko |
| author_sort | Hua Guo |
| collection | DOAJ |
| description | ABSTRACT Hundreds of sarbecoviruses have been found in bats, but only a fraction of them have the ability to infect cells using angiotensin-converting enzyme 2 (ACE2), the receptor for SARS-CoV and -2. To date, only ACE2-dependent sarbecoviruses have been isolated from field samples or grown in the laboratory. ACE2-independent sarbecoviruses, comprising the majority of the subgenus, have not been propagated in any type of cell culture, as the factors and conditions needed for their replication are completely unknown. Given the significant zoonotic threat posed by sarbecoviruses, cell culture models and in vitro tools are urgently needed to study the rest of this subgenus. We previously showed that the exogenous protease trypsin could facilitate cell entry of viral-like particles pseudotyped with spike protein from some of the ACE2-independent sarbecoviruses. Here, we tested if these conditions were sufficient to support bona fide viral replication using recombinant bat sarbecoviruses. In the presence of trypsin, some of the spike proteins from clade 2 viruses were capable of supporting bat sarbecovirus infection and replication in human and bat cells. Protease experiments showed a specific viral dependence on high levels of trypsin, as TMPRSS2 and furin had no effect on clade 2 virus entry. These results shed light on how sarbecoviruses transmit and coexist in their natural hosts, provide key insights for future efforts to isolate and grow these viruses from field samples, and further underscore the need for broadly protective, universal coronavirus vaccines. IMPORTANCE Our studies demonstrate that some unexplored sarbecoviruses are capable of replicating in human and bat cells in an ACE2-independent way but need a high trypsin environment. We found that trypsin is not compensated by other known proteases involved in some coronavirus entry. This work provides important information that the trypsin-dependent entry may be a widely employed mechanism for coronaviruses and will help for further understanding the biological features of the less-studied viruses. |
| first_indexed | 2024-04-12T00:55:48Z |
| format | Article |
| id | doaj.art-7386389ad9684d2eaff0dd771e8f5ee6 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-04-12T00:55:48Z |
| publishDate | 2022-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-7386389ad9684d2eaff0dd771e8f5ee62022-12-22T03:54:37ZengAmerican Society for MicrobiologymBio2150-75112022-12-0113610.1128/mbio.02566-22ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat CellsHua Guo0Ang Li1Tian-Yi Dong2Jia Su3Yu-Lin Yao4Yan Zhu5Zheng-Li Shi6Michael Letko7CAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, ChinaCAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, ChinaCAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, ChinaCAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, ChinaCAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, ChinaCAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, ChinaCAS Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, ChinaPaul G. Allen School for Global Health, Washington State University, Pullman, Washington, USAABSTRACT Hundreds of sarbecoviruses have been found in bats, but only a fraction of them have the ability to infect cells using angiotensin-converting enzyme 2 (ACE2), the receptor for SARS-CoV and -2. To date, only ACE2-dependent sarbecoviruses have been isolated from field samples or grown in the laboratory. ACE2-independent sarbecoviruses, comprising the majority of the subgenus, have not been propagated in any type of cell culture, as the factors and conditions needed for their replication are completely unknown. Given the significant zoonotic threat posed by sarbecoviruses, cell culture models and in vitro tools are urgently needed to study the rest of this subgenus. We previously showed that the exogenous protease trypsin could facilitate cell entry of viral-like particles pseudotyped with spike protein from some of the ACE2-independent sarbecoviruses. Here, we tested if these conditions were sufficient to support bona fide viral replication using recombinant bat sarbecoviruses. In the presence of trypsin, some of the spike proteins from clade 2 viruses were capable of supporting bat sarbecovirus infection and replication in human and bat cells. Protease experiments showed a specific viral dependence on high levels of trypsin, as TMPRSS2 and furin had no effect on clade 2 virus entry. These results shed light on how sarbecoviruses transmit and coexist in their natural hosts, provide key insights for future efforts to isolate and grow these viruses from field samples, and further underscore the need for broadly protective, universal coronavirus vaccines. IMPORTANCE Our studies demonstrate that some unexplored sarbecoviruses are capable of replicating in human and bat cells in an ACE2-independent way but need a high trypsin environment. We found that trypsin is not compensated by other known proteases involved in some coronavirus entry. This work provides important information that the trypsin-dependent entry may be a widely employed mechanism for coronaviruses and will help for further understanding the biological features of the less-studied viruses.https://journals.asm.org/doi/10.1128/mbio.02566-22sarbecovirusSARS-related coronavirusACE2-independenttrypsin-dependent |
| spellingShingle | Hua Guo Ang Li Tian-Yi Dong Jia Su Yu-Lin Yao Yan Zhu Zheng-Li Shi Michael Letko ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat Cells mBio sarbecovirus SARS-related coronavirus ACE2-independent trypsin-dependent |
| title | ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat Cells |
| title_full | ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat Cells |
| title_fullStr | ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat Cells |
| title_full_unstemmed | ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat Cells |
| title_short | ACE2-Independent Bat Sarbecovirus Entry and Replication in Human and Bat Cells |
| title_sort | ace2 independent bat sarbecovirus entry and replication in human and bat cells |
| topic | sarbecovirus SARS-related coronavirus ACE2-independent trypsin-dependent |
| url | https://journals.asm.org/doi/10.1128/mbio.02566-22 |
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