In:
PLOS Pathogens, Public Library of Science (PLoS), Vol. 19, No. 8 ( 2023-8-31), p. e1011614-
Abstract:
Despite unprecedented efforts, our therapeutic arsenal against SARS-CoV-2 remains limited. The conserved macrodomain 1 (Mac1) in NSP3 is an enzyme exhibiting ADP-ribosylhydrolase activity and a possible drug target. To determine the role of Mac1 catalytic activity in viral replication, we generated recombinant viruses and replicons encoding a catalytically inactive NSP3 Mac1 domain by mutating a critical asparagine in the active site. While substitution to alanine (N40A) reduced catalytic activity by ~10-fold, mutations to aspartic acid (N40D) reduced activity by ~100-fold relative to wild-type. Importantly, the N40A mutation rendered Mac1 unstable in vitro and lowered expression levels in bacterial and mammalian cells. When incorporated into SARS-CoV-2 molecular clones, the N40D mutant only modestly affected viral fitness in immortalized cell lines, but reduced viral replication in human airway organoids by 10-fold. In mice, the N40D mutant replicated at 〉 1000-fold lower levels compared to the wild-type virus while inducing a robust interferon response; all animals infected with the mutant virus survived infection. Our data validate the critical role of SARS-CoV-2 NSP3 Mac1 catalytic activity in viral replication and as a promising therapeutic target to develop antivirals.
Type of Medium:
Online Resource
ISSN:
1553-7374
DOI:
10.1371/journal.ppat.1011614
DOI:
10.1371/journal.ppat.1011614.g001
DOI:
10.1371/journal.ppat.1011614.g002
DOI:
10.1371/journal.ppat.1011614.g003
DOI:
10.1371/journal.ppat.1011614.g004
DOI:
10.1371/journal.ppat.1011614.s001
DOI:
10.1371/journal.ppat.1011614.s002
DOI:
10.1371/journal.ppat.1011614.s003
DOI:
10.1371/journal.ppat.1011614.s004
DOI:
10.1371/journal.ppat.1011614.s005
DOI:
10.1371/journal.ppat.1011614.s006
DOI:
10.1371/journal.ppat.1011614.s007
DOI:
10.1371/journal.ppat.1011614.s008
DOI:
10.1371/journal.ppat.1011614.s009
DOI:
10.1371/journal.ppat.1011614.s010
DOI:
10.1371/journal.ppat.1011614.s011
DOI:
10.1371/journal.ppat.1011614.s012
DOI:
10.1371/journal.ppat.1011614.s013
DOI:
10.1371/journal.ppat.1011614.r001
DOI:
10.1371/journal.ppat.1011614.r002
DOI:
10.1371/journal.ppat.1011614.r003
DOI:
10.1371/journal.ppat.1011614.r004
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2023
detail.hit.zdb_id:
2205412-1
Permalink