Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication
3CLpro is a key proteinase for SARS-CoV-2 replication and serves as an important target for antiviral drug development. However, how its activity is regulated intracellularly is still obscure. In this study, we developed a 3CLpro protease activity reporter system to examine the impact of various fac...
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Elsevier
2021
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oai:doaj.org-article:96766fb4cd0a4d358322855d4479906c2021-11-28T04:31:26ZOxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication2213-231710.1016/j.redox.2021.102199https://doaj.org/article/96766fb4cd0a4d358322855d4479906c2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2213231721003591https://doaj.org/toc/2213-23173CLpro is a key proteinase for SARS-CoV-2 replication and serves as an important target for antiviral drug development. However, how its activity is regulated intracellularly is still obscure. In this study, we developed a 3CLpro protease activity reporter system to examine the impact of various factors, including nutrient supplements, ions, pHs, or oxidative stress inducers, on 3CLpro protease activity. We found that oxidative stress could increase the overall activity of 3CLpro. Not altering the expression, oxidative stress decreased the solubility of 3CLpro in the lysis buffer containing 1% Triton-X-100. The Triton-X-100-insoluble 3CLpro was correlated with aggregates’ formation and responsible for the increased enzymatic activity. The disulfide bonds formed between Cys85 sites of 3CLpro protomers account for the insolubility and the aggregation of 3CLpro. Besides being regulated by oxidative stress, 3CLpro impaired the cellular antioxidant capacity by regulating the cleavage of GPx1 at its N-terminus. This cleavage could further elevate the 3CLpro-proximate oxidative activity, favor aggregation and activation of 3CLpro, and thus lead to a positive feedback loop. In summary, we reported that oxidative stress transforms 3CLpro into a detergent-insoluble form that is more enzymatically active, leading to increased viral replication/transcription. Our study provided mechanistic evidence that suggests the therapeutic potential of antioxidants in the clinical treatment of COVID-19 patients.Liubing DuYanchun XieKai ZhengNiu WangMingcheng GaoTing YuLiu CaoQianQian ShaoYong ZouWei XiaQianglin FangBo ZhaoDeyin GuoXiaoxue PengJi-An PanElsevierarticleSARS-CoV-23CLproOxidative stressInsolubleMedicine (General)R5-920Biology (General)QH301-705.5ENRedox Biology, Vol 48, Iss , Pp 102199- (2021) |
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| language |
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| topic |
SARS-CoV-2 3CLpro Oxidative stress Insoluble Medicine (General) R5-920 Biology (General) QH301-705.5 |
| spellingShingle |
SARS-CoV-2 3CLpro Oxidative stress Insoluble Medicine (General) R5-920 Biology (General) QH301-705.5 Liubing Du Yanchun Xie Kai Zheng Niu Wang Mingcheng Gao Ting Yu Liu Cao QianQian Shao Yong Zou Wei Xia Qianglin Fang Bo Zhao Deyin Guo Xiaoxue Peng Ji-An Pan Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication |
| description |
3CLpro is a key proteinase for SARS-CoV-2 replication and serves as an important target for antiviral drug development. However, how its activity is regulated intracellularly is still obscure. In this study, we developed a 3CLpro protease activity reporter system to examine the impact of various factors, including nutrient supplements, ions, pHs, or oxidative stress inducers, on 3CLpro protease activity. We found that oxidative stress could increase the overall activity of 3CLpro. Not altering the expression, oxidative stress decreased the solubility of 3CLpro in the lysis buffer containing 1% Triton-X-100. The Triton-X-100-insoluble 3CLpro was correlated with aggregates’ formation and responsible for the increased enzymatic activity. The disulfide bonds formed between Cys85 sites of 3CLpro protomers account for the insolubility and the aggregation of 3CLpro. Besides being regulated by oxidative stress, 3CLpro impaired the cellular antioxidant capacity by regulating the cleavage of GPx1 at its N-terminus. This cleavage could further elevate the 3CLpro-proximate oxidative activity, favor aggregation and activation of 3CLpro, and thus lead to a positive feedback loop. In summary, we reported that oxidative stress transforms 3CLpro into a detergent-insoluble form that is more enzymatically active, leading to increased viral replication/transcription. Our study provided mechanistic evidence that suggests the therapeutic potential of antioxidants in the clinical treatment of COVID-19 patients. |
| format |
article |
| author |
Liubing Du Yanchun Xie Kai Zheng Niu Wang Mingcheng Gao Ting Yu Liu Cao QianQian Shao Yong Zou Wei Xia Qianglin Fang Bo Zhao Deyin Guo Xiaoxue Peng Ji-An Pan |
| author_facet |
Liubing Du Yanchun Xie Kai Zheng Niu Wang Mingcheng Gao Ting Yu Liu Cao QianQian Shao Yong Zou Wei Xia Qianglin Fang Bo Zhao Deyin Guo Xiaoxue Peng Ji-An Pan |
| author_sort |
Liubing Du |
| title |
Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication |
| title_short |
Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication |
| title_full |
Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication |
| title_fullStr |
Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication |
| title_full_unstemmed |
Oxidative stress transforms 3CLpro into an insoluble and more active form to promote SARS-CoV-2 replication |
| title_sort |
oxidative stress transforms 3clpro into an insoluble and more active form to promote sars-cov-2 replication |
| publisher |
Elsevier |
| publishDate |
2021 |
| url |
https://doaj.org/article/96766fb4cd0a4d358322855d4479906c |
| work_keys_str_mv |
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