Early cysteine-dependent inactivation of 26S proteasomes does not involve particle disassembly

Hugo, Martín; Korovila, Ioanna; Köhler, M; García-García, Carlos; Cabrera-García, J. Daniel; Marina, Anabel; Martínez‐Ruiz, Antonio; Grune, Tilman

doi:10.1016/j.redox.2018.02.016

Cited by 6 publications

(2 citation statements)

References 24 publications

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“…Disruption of proteostasis can result in the deposition of protein aggregates and be harmful for cell survival [45,46]. Protein degradation by the proteasome changes with aging and is regulated by redox modulation, especially thiol oxidation [47,48]. Here, we identified three age-oxidized cysteines in proteasome 26S subunits and one age-reduced cysteine in heat shock protein 70 (HSP70), providing further evidence for the idea that the 26S proteasome is vulnerable to redox regulation, and that its chaperone HSP70 responds to redox regulation [49,50].…”

Section: Discussionmentioning

confidence: 99%

Redox States of Protein Cysteines in Pathways of Protein Turnover and Cytoskeleton Dynamics Are Changed with Aging and Reversed by Slc7a11 Restoration in Mouse Lung Fibroblasts

Zheng

Merchant

Burke

et al. 2020

Oxidative Medicine and Cellular Longevity

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Slc7a11 is the key component of system Xc-, an antiporter that imports cystine (CySS) and exports glutamate. It plays an important role in cellular defense against oxidative stress because cysteine (Cys), reduced from CySS, is used for and limits the synthesis of glutathione (GSH). We have shown that downregulation of Slc7a11 is responsible for oxidation of extracellular Cys/CySS redox potential in lung fibroblasts from old mice. However, how age-related change of Slc7a11 expression affects the intracellular redox environment of mouse lung fibroblasts remains unexplored. The purpose of this study is to evaluate the effects of aging on the redox states of intracellular proteins and to examine whether Slc7a11 contributes to the age-dependent effects. Iodoacetyl Tandem Mass Tags were used to differentially label reduced and oxidized forms of Cys residues in primary lung fibroblasts from young and old mice, as well as old fibroblasts transfected with Slc7a11. The ratio of oxidized/reduced forms (i.e., redox state) of a Cys residue was determined via multiplexed tandem mass spectrometry. Redox states of 151 proteins were different in old fibroblasts compared to young fibroblasts. Slc7a11 overexpression restored redox states of 104 (69%) of these proteins. Ingenuity Pathway Analysis (IPA) showed that age-dependent Slc7a11-responsive proteins were involved in pathways of protein translation initiation, ubiquitin-proteasome-mediated degradation, and integrin-cytoskeleton-associated signaling. Gene ontology analysis showed cell adhesion, protein translation, and organization of actin cytoskeleton were among the top enriched terms for biological process. Protein-protein interaction network demonstrated the interactions between components of the three enriched pathways predicted by IPA. Follow-up experiments confirmed that proteasome activity was lower in old cells than in young cells and that upregulation of Slc7a11 expression by sulforaphane restored this activity. This study finds that aging results in changes of redox states of proteins involved in protein turnover and cytoskeleton dynamics, and that upregulating Slc7a11 can partially restore the redox states of these proteins.

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Section: Discussionmentioning

confidence: 99%

Redox States of Protein Cysteines in Pathways of Protein Turnover and Cytoskeleton Dynamics Are Changed with Aging and Reversed by Slc7a11 Restoration in Mouse Lung Fibroblasts

Zheng

Merchant

Burke

et al. 2020

Oxidative Medicine and Cellular Longevity

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“…According to the second assumption, the decreased Gpx1 is due to their antioxidant activity for scavenging excessive ROS generated in the mitotic phase (Lubos et al 2011;Hugo et al 2018).…”

Section: Discussionmentioning

confidence: 99%

Association of Gpx1 fluctuation in cell cycle progression

Aktar

Kafi

Dahiya

2019

In Vitro Cell.Dev.Biol.-Animal

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This research demonstrates fluctuation of glutathione peroxidase1 (Gpx1) throughout cell cycle progression with significant decreased expression at mitosis of HeLa cell. This was achieved with western blot (WB) analysis of target proteins from each phase of synchronized cells. The synchronizations were performed with double thymidine (T/T) for G1/S arrest and thymidine followed by nocodazol (T/N) for G2/M arrest. The G1/S arrested cells were released in fresh medium for 3, 6, 9, 10 and 15h to obtain cell at each phase such as Gap1 (G1), synthesis (S), gap2 (G2), mitosis (M) and gap1 (G1) phase, respectively for investigating Gpx1 expression throughout a complete cycle. The synchronizations were confirmed using fluorescence activated cell sorting (FACS) and WB analysis of phase specific markers. The fluctuations of Gpx1 expression were verified with universal protein actin and peroxiredoxin 1 (Prx1) which are stable throughout the cell cycle. Intriguingly, immunoblots showed the level of Gpx1 decreases at mitosis phase and increased during mitotic exit to G1 phase in HeLa cells, while Prx1 protein level remained constant. The fractionation experiments reveal that only the cytosolic Gpx1 was decreased while their levels at mitochondria remain constant. The heighest levels of mitochondrial ROS were measured in mitosis phase with FACS analysis using Mito sox indicating that antioxidant activity of Gpx1 for detoxifying excessive induced endogenous reactive oxygen species (ROS) in the mitosis phase could be the reason for such decreasing level.For unfolding the molecular mechanism of such decreased expression, the Gpx1 was investigated at transcriptional, translational and proteosomal level. The results revealed that translational mechanism is involve in the decreased expression rather than transcriptional or proteosomal degradation at mitosis phase. This finding supports that Gpx1 involved in the cell cycle progression through regulation of endogenous ROS. Based on this observation further research could uncover their possible association with the infinitive division of a cancer cell.

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Endogenous anti-tumorigenic nitro-fatty acids inhibit the ubiquitin-proteasome system by directly targeting the 26S proteasome

Brat

Phuoc

Awad

et al. 2023

Cell Chemical Biology

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Early cysteine-dependent inactivation of 26S proteasomes does not involve particle disassembly

Cited by 6 publications

References 24 publications

Redox States of Protein Cysteines in Pathways of Protein Turnover and Cytoskeleton Dynamics Are Changed with Aging and Reversed by Slc7a11 Restoration in Mouse Lung Fibroblasts

Redox States of Protein Cysteines in Pathways of Protein Turnover and Cytoskeleton Dynamics Are Changed with Aging and Reversed by Slc7a11 Restoration in Mouse Lung Fibroblasts

Association of Gpx1 fluctuation in cell cycle progression

Endogenous anti-tumorigenic nitro-fatty acids inhibit the ubiquitin-proteasome system by directly targeting the 26S proteasome

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