<p>Phylogenetic and geographic distribution of bat sarbecoviruses and their bat ACE2 tropism.</p>

Offset images highlight the conserved amino acid residues across the different bat ACE2s relative to <i>Rh. affinis</i> ACE2, and the conservation of bat sarbecovirus RBDs compared to SARS-CoV-2, screened in this study. The ACE2-RBD interacting residues are denoted. <b>(D)</b> Maximum likelihood phylogeny of the bat sarbecoviruses full-length Spike and bat ACE2 amino acid sequences are shown. Bat sarbecovirus Spikes were used to generate lentiviral-based pseudotypes and used to infect BHK-21 cells overexpressing different bat ACE2s or human ACE2.

A heatmap illustrating receptor usage is shown, representing the mean log RLU of 3−5 separate experiments. A vector-only control (pDISPLAY) was included to demonstrate specificity and to set the background signal. The cognate bat receptor for each virus, where possible is highlighted with a red box.

Absence of a red square is indicative of the lack of cognate receptor from our screen, either due to unavailability of ACE2 sequence, or because the original bat host is yet unknown (RhGB07: <i>Rh. hipposideros</i>; SARS-CoV-2, SARS-CoV-1: unknown; BANAL-20-52: <i>Rh. malayanus</i>; BANAL-20-236: <i>Rh. marshalli</i>; RacCS203 – <i>Rh. acuminatus</i>). <b>(E)</b> Sequence alignment of bat sarbecovirus RBDs at the ACE2-binding interface, with key amino acid residues denoted, using SARS-CoV-2 numbering.

Regions of amino acid deletions between sequences are denoted in red. The corresponding human and <i>Rh.affinis</i> ACE2 residues that bind SARS-CoV-2 RBD are highlighted, with black indicating amino acid residues that are the same between human ACE2 and <i>Rh. affinis</i> ACE2, green indicating the same residues but a different amino acid, and the remainder highlighting unique binding residues for human ACE2 (blue) or <i>Rh. affinis</i> ACE2 (pink) only.

The data underlying this Figure can be found in <a href="plosbiology.org/article/info:doi/10.1371/journal.pbio.3003944#pbio.3003944.s017" target="_blank">S1 Data</a>.</p>