Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation

Mutp53 level is widely variable among individual cancer cells in tumor tissues, and within cells a higher level of mutp53 is usually observed in the nucleus as compared to the cytoplasm. This spatial heterogeneity in mutp53 expression has been well documented and likely plays an important role in tu...

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Main Authors: Liansheng Wang, Suqin Zhong, Xinru Fan, Yuxue Xu, Meimei Wang, Youcui Xu, Yuanyuan Cai, Zhong Cao, Zhiming Ye, Longping Wen, Pengfei Wei
Format: Article
Language:English
Published: Elsevier 2025-04-01
Series:Redox Biology
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Online Access:http://www.sciencedirect.com/science/article/pii/S2213231725000357
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author Liansheng Wang
Suqin Zhong
Xinru Fan
Yuxue Xu
Meimei Wang
Youcui Xu
Yuanyuan Cai
Zhong Cao
Zhiming Ye
Longping Wen
Pengfei Wei
author_facet Liansheng Wang
Suqin Zhong
Xinru Fan
Yuxue Xu
Meimei Wang
Youcui Xu
Yuanyuan Cai
Zhong Cao
Zhiming Ye
Longping Wen
Pengfei Wei
author_sort Liansheng Wang
collection DOAJ
description Mutp53 level is widely variable among individual cancer cells in tumor tissues, and within cells a higher level of mutp53 is usually observed in the nucleus as compared to the cytoplasm. This spatial heterogeneity in mutp53 expression has been well documented and likely plays an important role in tumor therapeutic resistance. However, its underlying mechanism remains poorly understood.In this study, we first revealed a critical role of micro-environmental reducing status in regulating mutp53 stability and spatially heterogeneous accumulation. Immunofluorescence and ThiolTracker Violet dye staining demonstrated a clear correlation between the cellular mutp53 level and the reducibility in the patient-derived tumor tissues and mutp53-expressing cancer cell lines. The nucleus exhibited both higher reducibility and more mutp53 accumulation than the cytoplasm did. Supplementing GSH exacerbated the accumulation of mutp53, while consuming GSH led to extensive depletion of mutp53, suggesting that the environmental reducing status kept mutp53 stability. Mechanistically, S-glutathionylation could trigger ubiquitination and proteasomal degradation of mutp53. A highly-reducing local environment preserved mutp53 stability by inhibiting glutathionylation and subsequent proteasomal degradation of mutp53, which also provided an explanation for the differential accumulation of mutp53 proteins in the nucleus and cytoplasm. Thirdly, we revealed that the expression level of glutathione reductase (GR) was positively correlated with mutp53 accumulation across the cultured mutp53-expressing cell lines, patient-derived tumor tissues and patient databases. Over-expression of GR reinforced the environmental reducibility, affected glutathionylation and improved mutp53 accumulation, while inhibiting GR either by chemical inhibitors or genetic approach induced massive clearance of a variety of mutp53 and effectively retarded the growth of p53-mutated cell-derived xenografts in mice.These studies provided an explanation for the widely-observed spatial heterogeneous accumulation of mutp53 proteins, and inhibiting GR or directly consuming GSH represented a promising strategy for mutp53 carrying cancer therapy.
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spelling doaj-art-3af10764ffb84bc68b6fbdc311badf782025-08-20T03:02:56ZengElsevierRedox Biology2213-23172025-04-018110352210.1016/j.redox.2025.103522Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylationLiansheng Wang0Suqin Zhong1Xinru Fan2Yuxue Xu3Meimei Wang4Youcui Xu5Yuanyuan Cai6Zhong Cao7Zhiming Ye8Longping Wen9Pengfei Wei10Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, ChinaGuangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, China; School of Medicine & Institute for Life Sciences, South China University of Technology, Guangzhou, 510006, ChinaSchool of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, 264003, ChinaSchool of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, 264003, China; Yantai Stem Cell and Regenerative Medicine Key Laboratory, Binzhou Medical University, Yantai, 264003, ChinaSchool of Medicine & Institute for Life Sciences, South China University of Technology, Guangzhou, 510006, ChinaGuangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, ChinaSchool of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, 264003, ChinaSchool of Biomedical Engineering, Shenzhen Campus of Sun Yat-sen University, Shenzhen, Guangdong, 518107, China; Shenzhen International Institute for Biomedical Research, Silver Star Hi-tech Park, Longhua District, Shenzhen, Guangdong, 518116, ChinaGuangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, China; School of Medicine & Institute for Life Sciences, South China University of Technology, Guangzhou, 510006, China; Corresponding author. Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, China.Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, China; Corresponding author.School of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, 264003, China; Corresponding author. School of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, 264003, China.Mutp53 level is widely variable among individual cancer cells in tumor tissues, and within cells a higher level of mutp53 is usually observed in the nucleus as compared to the cytoplasm. This spatial heterogeneity in mutp53 expression has been well documented and likely plays an important role in tumor therapeutic resistance. However, its underlying mechanism remains poorly understood.In this study, we first revealed a critical role of micro-environmental reducing status in regulating mutp53 stability and spatially heterogeneous accumulation. Immunofluorescence and ThiolTracker Violet dye staining demonstrated a clear correlation between the cellular mutp53 level and the reducibility in the patient-derived tumor tissues and mutp53-expressing cancer cell lines. The nucleus exhibited both higher reducibility and more mutp53 accumulation than the cytoplasm did. Supplementing GSH exacerbated the accumulation of mutp53, while consuming GSH led to extensive depletion of mutp53, suggesting that the environmental reducing status kept mutp53 stability. Mechanistically, S-glutathionylation could trigger ubiquitination and proteasomal degradation of mutp53. A highly-reducing local environment preserved mutp53 stability by inhibiting glutathionylation and subsequent proteasomal degradation of mutp53, which also provided an explanation for the differential accumulation of mutp53 proteins in the nucleus and cytoplasm. Thirdly, we revealed that the expression level of glutathione reductase (GR) was positively correlated with mutp53 accumulation across the cultured mutp53-expressing cell lines, patient-derived tumor tissues and patient databases. Over-expression of GR reinforced the environmental reducibility, affected glutathionylation and improved mutp53 accumulation, while inhibiting GR either by chemical inhibitors or genetic approach induced massive clearance of a variety of mutp53 and effectively retarded the growth of p53-mutated cell-derived xenografts in mice.These studies provided an explanation for the widely-observed spatial heterogeneous accumulation of mutp53 proteins, and inhibiting GR or directly consuming GSH represented a promising strategy for mutp53 carrying cancer therapy.http://www.sciencedirect.com/science/article/pii/S2213231725000357Mutant p53GlutathionylationGlutathione reductaseGSH/GSSG ratioUbiquitin-proteasome systemMutant p53-carrying cancer
spellingShingle Liansheng Wang
Suqin Zhong
Xinru Fan
Yuxue Xu
Meimei Wang
Youcui Xu
Yuanyuan Cai
Zhong Cao
Zhiming Ye
Longping Wen
Pengfei Wei
Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation
Redox Biology
Mutant p53
Glutathionylation
Glutathione reductase
GSH/GSSG ratio
Ubiquitin-proteasome system
Mutant p53-carrying cancer
title Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation
title_full Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation
title_fullStr Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation
title_full_unstemmed Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation
title_short Glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation
title_sort glutathione reductase underlies the stability of mutant p53 by antagonizing protein glutathionylation
topic Mutant p53
Glutathionylation
Glutathione reductase
GSH/GSSG ratio
Ubiquitin-proteasome system
Mutant p53-carrying cancer
url http://www.sciencedirect.com/science/article/pii/S2213231725000357
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