Cell entry of SARS-CoV-2 conferred by angiotensin-converting enzyme 2 (ACE2) of different species

Abstract

1The outbreak of the severe acute respiratory syndrome coronavirus 2 2 (SARS-CoV-2) poses a huge threat to many countries around the world. However, 3 where is it origin and which animals are sensitive to cross-species transmission is 4 unclear. The interaction of virus and cell receptor is a key determinant of host range 5 for the novel coronavirus. Angiotensin-converting enzyme 2 (ACE2) is demonstrated 6 as the primary entry receptor for SARS-CoV-2. In this study, we evaluated the 7 SARS-CoV-2 entry mediated by ACE2 of 11 different species of animals, and 8 discovered that ACE2 of Rhinolophus sinicus (Chinese horseshoe bat), Felis catus 9 (domestic cat), Canis lupus familiaris (dog), Sus scrofa (pig), Capra hircus (goat) and 10 especially Manis javanica (Malayan pangolin) were able to render SARS-CoV-2 entry 11 in non-susceptible cells. This is the first report that ACE2 of Pangolin could mediate 12 SARS-CoV-2 entry which increases the presume that SARS-CoV-2 may have a 13 pangolin origin. However, none of the ACE2 proteins from Rhinolophus 14 ferrumequinum (greater horseshoe bat), Gallus gallus (chicken), Notechis scutatus 15 (mainland tiger snake), Mus musculus (house mouse) rendered SARS-CoV-2 entry. 16 Specifically, a natural isoform of Macaca mulatta (Rhesus monkey) ACE2 with a 17 mutation of Y217N was resistance to infection, which rises the possible impact of this 18 type of ACE2 during monkey studies of SARS-CoV-2. Overall, these results clarify 19 that SARS-CoV-2 could engage receptors of multiple species of animals and it is a 20 perplexed work to track SARS-CoV-2 origin and its intermediate hosts. 21 22 IMPORTANCE 1In this study, we illustrated that SARS-CoV-2 is able to engage receptors of 2 multiple species of animals. This indicated that it may be a perplexed work to track 3 SARS-CoV-2 origin and discover its intermediate hosts. This feature of virus is 4 considered to potentiate its diverse cross-species transmissibility. Of note, here is the 5 first report that ACE2 of Pangolin could mediate SARS-CoV-2 entry which increases 6 the possibility that SARS-CoV-2 may have a pangolin origin. And we also 7 demonstrated that not all species of bat were sensitive to SARS-CoV-2 infection. At 8 last, it is also important to detect the expression ratio of the Y217N ACE2 to the 9 prototype in Rhesus monkeys to be recruited for studies on SARS-CoV-2 infection.

Multimerization- and glycosylation-dependent receptor binding of SARS-CoV-2 spike proteins

“…However, these analyses are limited as it does not determine expression biochemically on epithelial cells that make contact to the outside world [29]. Several groups have transfected variants of ACE2 in cells to analyze transduction by SARS-CoV-pseudo viruses, although informative these studies do not provide information on the natural expression pattern of ACE2 in susceptible hosts [30, 31]. A detailed understanding of the difference between animal species and cell-specific expression of ACE2 at the molecular level is essential, as this can provide valuable knowledge on potential hosts that can be susceptible to SARS-like coronaviruses.…”

The Potential Intermediate Hosts for SARS-CoV-2

“… Tang et al (2020) conducted virus infectivity studies using HEK293T cells expressing ACE2 from 11 species of animals, and found that pangolin-ACE2 could mediate SARS-CoV-2 entry. Besides, ACE2 of Rhinolophus ferrumequinum cannot support the entry of SARS-CoV-2 that is a capability of ACE2 of Rhinolophus sinicus in experiments conducted by Tang et al (2020) and Zhou P. et al (2020) .…”

“… Tang et al (2020) conducted virus infectivity studies using HEK293T cells expressing ACE2 from 11 species of animals, and found that pangolin-ACE2 could mediate SARS-CoV-2 entry. Besides, ACE2 of Rhinolophus ferrumequinum cannot support the entry of SARS-CoV-2 that is a capability of ACE2 of Rhinolophus sinicus in experiments conducted by Tang et al (2020) and Zhou P. et al (2020) . By using pseudotyping particles of SARS-CoV-2 Spike mimics particle entry and quantitative cell-cell fusion assay, Conceicao et al (2020) observed that pangolin-ACE2 sustained higher levels of entry than hACE2.…”

Host Diversity and Potential Transmission Pathways of SARS-CoV-2 at the Human-Animal Interface

“…Furthermore, N-glycosylation motifs (e.g., N53) are conserved within all species and could potentially attach to the S-protein [ 54 ]. Based on protein modeling approaches, extensive vertebrate diversity has been reported as potentially susceptible to SARS-CoV-2 [ 122 , 125 ]. However, further experimental data will need to be conducted to confirm the results of the ACE-2 sequencing data [ 122 ].…”

“…Moreover, the ACE-2 receptors of pigs could potentially be used by SARS-CoV-2 [ 277 ]. It is speculated that pig ACE-2 receptors could genetically recombine to mediate SARS-CoV-2 entry into other species similar to other swine recombinant events [ 125 ].…”