Using cells derived from Eptesicus fuscus (big brown bats) expressing human ACE2 to mimic infection in SARS-CoV-2′s zoonotic reservoir (12), we showed that the R203M and T205I mutations all reduced replication and SARS-CoV-2 N phosphorylation relative to wildtype virus, while the KR mutant grew to equal titer. Here, ACE2 is linked to infection.