Recombinant SARS-CoV-2 NSP15 Protein

Nsp15-like endoribonucleases are a characteristic of all coronavirus family members. Biochemical experiments with recombinant Nsp15 have established that it preferentially cleaves RNA substrates 3′ of uridines, and therefore Nsp15 is commonly called endoU alluding to its cleavage specificity. Conservation of Nsp15 across Coronaviridae suggests that its endonuclease function is critical for their viral life cycle, however, the specific role of Nsp15 in viral propagation is still unclear. Nsp15 was initially thought to play an essential proofreading role in viral replication until it was shown that the coronavirus, mouse hepatitis virus (MHV), can replicate with a catalytically deficient variant of Nsp15 in cell culture. More recent work suggests that rather than functioning in viral RNA synthesis, Nsp15 nuclease activity is important to evade activation of host immune responses. Recent analysis of viral RNA from MHV-infected cells harboring a catalytically deficient Nsp15 revealed an accumulation of 12–17 polyuridine tracts at the 5′-end of the negative-strand viral RNA intermediates. Considering that polyuridine negative-strand RNA elicits an interferon-mediated response, this suggests a role for Nsp15 in regulating the length of polyuridines found at the 5′-end of negative-strand viral RNA to evade activation of the host innate immune response.