Literature detail

Expanded ACE2 dependencies of diverse SARS-like coronavirus receptor binding domains.

Sarah M Roelle¹ Nidhi Shukla¹ Anh T Pham¹ Anna M Bruchez¹ Kenneth A Matreyek¹

Affiliations 1 institutions

Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, Ohio, United States of America.

PMID 35895696 2022 PLoS Biol eng epublish

Article

Publication summary

Viral spillover from animal reservoirs can trigger public health crises and cripple the world economy. Knowing which viruses are primed for zoonotic transmission can focus surveillance efforts and mitigation strategies for future pandemics. Successful engagement of receptor protein orthologs is necessary during cross-species transmission. The clade 1 sarbecoviruses including Severe Acute Respiratory Syndrome-related Coronavirus (SARS-CoV) and SARS-CoV-2 enter cells via engagement of angiotensin converting enzyme-2 (ACE2), while the receptor for clade 2 and clade 3 remains largely uncharacterized. We developed a mixed cell pseudotyped virus infection assay to determine whether various clades 2 and 3 sarbecovirus spike proteins can enter HEK 293T cells expressing human or Rhinolophus horseshoe bat ACE2 proteins. The receptor binding domains from BtKY72 and Khosta-2 used human ACE2 for entry, while BtKY72 and Khosta-1 exhibited widespread use of diverse rhinolophid ACE2s. A lysine at ACE2 position 31 appeared to be a major determinant of the inability of these RBDs to use a certain ACE2 sequence. The ACE2 protein from Rhinolophus alcyone engaged all known clade 3 and clade 1 receptor binding domains. We observed little use of Rhinolophus ACE2 orthologs by the clade 2 viruses, supporting the likely use of a separate, unknown receptor. Our results suggest that clade 3 sarbecoviruses from Africa and Europe use Rhinolophus ACE2 for entry, and their spike proteins appear primed to contribute to zoonosis under the right conditions.

Angiotensin-Converting Enzyme 2 Chiroptera COVID-19 Receptors, Coronavirus Animals Humans Peptidyl-Dipeptidase A Protein Binding Receptors, Virus SARS-CoV-2 Spike Glycoprotein, Coronavirus spike protein, SARS-CoV-2

Structured evidence records

Evidence records

11 total

Receptor Usage 5 records

Receptor Usage Extraction confidence 1.00

Key finding

BtKY72 and Khosta-2 RBDs can use human ACE2, whereas BtKY72 and Khosta-1 utilize various bat ACE2 orthologs for viral entry.

Virus

Host

Location

Supporting text

The receptor binding domains from BtKY72 and Khosta-2 used human ACE2 for entry, while BtKY72 and Khosta-1 exhibited widespread use of diverse rhinolophid ACE2s.

Method: pseudotyped virus infection assay
Receptors: ACE2

Receptor Usage Extraction confidence 1.00

Key finding

BtKY72 and Khosta-1 RBDs interact with multiple Rhinolophus bat ACE2 orthologs for receptor-mediated entry.

Virus

Host

Location

Supporting text

BtKY72 and Khosta-1 exhibited widespread use of diverse rhinolophid ACE2s.

Method: pseudotyped virus infection assay
Receptors: ACE2

Receptor Usage Extraction confidence 1.00

Key finding

Clade 2 sarbecoviruses do not efficiently use Rhinolophus bat ACE2, suggesting an alternative receptor mediates entry.

Virus

Host

Location

Supporting text

We observed little use of Rhinolophus ACE2 orthologs by the clade 2 viruses, supporting the likely use of a separate, unknown receptor.

Method: pseudotyped virus infection assay
Receptors: ACE2

Receptor Usage Extraction confidence 1.00

Key finding

Rhinolophus alcyone ACE2 is compatible with clade 1 and clade 3 sarbecovirus RBDs.

Virus

Host

Location

Supporting text

The ACE2 protein from Rhinolophus alcyone engaged all known clade 3 and clade 1 receptor binding domains.

Method: pseudotyped virus infection assay
Receptors: ACE2

Receptor Usage Extraction confidence 1.00

Key finding

Rhinolophus alcyone ACE2 binds clade 3 sarbecovirus RBDs enabling receptor-mediated entry.

Virus

Host

Location

Supporting text

The ACE2 protein from Rhinolophus alcyone engaged all known clade 3 and clade 1 receptor binding domains.

Method: pseudotyped virus infection assay
Receptors: ACE2

Host Range Experiment 3 records

Host Range Experiment Extraction confidence 0.95

Key finding

BtKY72 and Khosta-2 receptor binding domains were experimentally shown to mediate entry via human ACE2 using a pseudotyped virus infection assay.

Virus

Host

Location

Supporting text

We developed a mixed cell pseudotyped virus infection assay to determine whether various clades 2 and 3 sarbecovirus spike proteins can enter HEK 293T cells expressing human or Rhinolophus horseshoe bat ACE2 proteins. The receptor binding domains from BtKY72 and Khosta-2 used human ACE2 for entry.

Method: pseudotyped virus infection assay
Experimental system: pseudovirus assay

Host Range Experiment Extraction confidence 0.95

Key finding

BtKY72 and Khosta-1 receptor binding domains entered cells expressing multiple Rhinolophus bat ACE2 proteins in a pseudovirus infection assay.

Virus

Host

Location

Supporting text

We developed a mixed cell pseudotyped virus infection assay to determine whether various clades 2 and 3 sarbecovirus spike proteins can enter HEK 293T cells expressing human or Rhinolophus horseshoe bat ACE2 proteins. BtKY72 and Khosta-1 exhibited widespread use of diverse rhinolophid ACE2s.

Method: pseudotyped virus infection assay
Experimental system: pseudovirus assay

Host Range Experiment Extraction confidence 0.95

Key finding

Rhinolophus alcyone ACE2 mediated entry of all known clade 3 and clade 1 sarbecovirus receptor binding domains in pseudovirus assays.

Virus

Host

Location

Supporting text

We developed a mixed cell pseudotyped virus infection assay to determine whether various clades 2 and 3 sarbecovirus spike proteins can enter HEK 293T cells expressing human or Rhinolophus horseshoe bat ACE2 proteins. The ACE2 protein from Rhinolophus alcyone engaged all known clade 3 and clade 1 receptor binding domains.

Method: pseudotyped virus infection assay
Experimental system: pseudovirus assay

Molecular Adaptation 3 records

Molecular Adaptation Extraction confidence 0.80

Key finding

Spike receptor binding domains from BtKY72 and Khosta-2 sarbecoviruses can engage human ACE2, indicating molecular adaptation of these viruses to human receptor usage.

Virus

Host

Location

Supporting text

The receptor binding domains from BtKY72 and Khosta-2 used human ACE2 for entry, while BtKY72 and Khosta-1 exhibited widespread use of diverse rhinolophid ACE2s.

Genes or proteins: spike; receptor binding domain
Receptors: ACE2
Mechanism types: receptor_binding; cell_entry

Molecular Adaptation Extraction confidence 0.80

Key finding

Spike receptor binding domains from BtKY72 and Khosta-1 sarbecoviruses can engage multiple Rhinolophus ACE2 orthologs, showing expanded receptor usage across bat species.

Virus

Host

Location

Supporting text

BtKY72 and Khosta-1 exhibited widespread use of diverse rhinolophid ACE2s.

Genes or proteins: spike; receptor binding domain
Receptors: ACE2
Mechanism types: receptor_binding; cell_entry

Molecular Adaptation Extraction confidence 0.80

Key finding

A lysine at ACE2 residue 31 was identified as a key molecular determinant affecting receptor usage by sarbecovirus RBDs.

Virus

Host

Location

Supporting text

A lysine at ACE2 position 31 appeared to be a major determinant of the inability of these RBDs to use a certain ACE2 sequence.

Genes or proteins: ACE2
Receptors: ACE2
Mutations: K31
Mechanism types: receptor_binding; host_factor_interaction

Citation context

References

71 references

Reference network

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Evidence overview

Evidence 11

Types 3

Virus 2

Host 7

Evidence types

Receptor Usage 5 Host Range Experiment 3 Molecular Adaptation 3

Linked entities

Viruses

Hosts

Locations

Publication metadata

Journal: PLoS biology
Volume / Issue / Pages: 20 / 7 / e3001738
ISSN: 1545-7885
Journal country: United States
DOI: 10.1371/journal.pbio.3001738