Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) non-structural protein 5 (nsp5) is a cysteine protease involved in viral replication and suppression of the host immune system. The substrate-binding domain of nsp5 is important for its protease activity. However, the relationship between...

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Main Authors: Yoshiro Sugiura, Kenta Shimizu, Tatsuki Takahashi, Shiori Ueno, Haruka Tanigou, Sodbayasgalan Amarbayasgalan, Wataru Kamitani
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2024-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0312800
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author Yoshiro Sugiura
Kenta Shimizu
Tatsuki Takahashi
Shiori Ueno
Haruka Tanigou
Sodbayasgalan Amarbayasgalan
Wataru Kamitani
author_facet Yoshiro Sugiura
Kenta Shimizu
Tatsuki Takahashi
Shiori Ueno
Haruka Tanigou
Sodbayasgalan Amarbayasgalan
Wataru Kamitani
author_sort Yoshiro Sugiura
collection DOAJ
description Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) non-structural protein 5 (nsp5) is a cysteine protease involved in viral replication and suppression of the host immune system. The substrate-binding domain of nsp5 is important for its protease activity. However, the relationship between nsp5 protease activity and viral replication remains unclear. We confirmed the importance of amino acid T25 in the nsp5 substrate-binding domain for viral replication using a split luciferase assay. By generating recombinant viruses using bacterial artificial chromosomes, we found that the proliferation of viruses with the T25I mutation in nsp5 was cell-dependent in culture. Furthermore, mice infected with the T25I mutant recombinant virus with a mouse acclimation backbone showed weight loss and increased lung viral load, similar to the wild-type (WT) infected group, up to 3 days after infection. However, after day 4, the lung viral load was significantly reduced in the T25I-infected group compared to that in the WT-infected group. This suggests that nsp5 T25 is involved in the pathogenesis of SARS-CoV-2.
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institution Kabale University
issn 1932-6203
language English
publishDate 2024-01-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS ONE
spelling doaj-art-4b3ff1c5080c457f8b5018fbba81be812024-12-10T05:32:07ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-011912e031280010.1371/journal.pone.0312800Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.Yoshiro SugiuraKenta ShimizuTatsuki TakahashiShiori UenoHaruka TanigouSodbayasgalan AmarbayasgalanWataru KamitaniSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) non-structural protein 5 (nsp5) is a cysteine protease involved in viral replication and suppression of the host immune system. The substrate-binding domain of nsp5 is important for its protease activity. However, the relationship between nsp5 protease activity and viral replication remains unclear. We confirmed the importance of amino acid T25 in the nsp5 substrate-binding domain for viral replication using a split luciferase assay. By generating recombinant viruses using bacterial artificial chromosomes, we found that the proliferation of viruses with the T25I mutation in nsp5 was cell-dependent in culture. Furthermore, mice infected with the T25I mutant recombinant virus with a mouse acclimation backbone showed weight loss and increased lung viral load, similar to the wild-type (WT) infected group, up to 3 days after infection. However, after day 4, the lung viral load was significantly reduced in the T25I-infected group compared to that in the WT-infected group. This suggests that nsp5 T25 is involved in the pathogenesis of SARS-CoV-2.https://doi.org/10.1371/journal.pone.0312800
spellingShingle Yoshiro Sugiura
Kenta Shimizu
Tatsuki Takahashi
Shiori Ueno
Haruka Tanigou
Sodbayasgalan Amarbayasgalan
Wataru Kamitani
Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.
PLoS ONE
title Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.
title_full Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.
title_fullStr Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.
title_full_unstemmed Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.
title_short Amino acid T25 in the substrate-binding domain of SARS-CoV-2 nsp5 is involved in viral replication in the mouse lung.
title_sort amino acid t25 in the substrate binding domain of sars cov 2 nsp5 is involved in viral replication in the mouse lung
url https://doi.org/10.1371/journal.pone.0312800
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