E3 ubiquitin ligases STUB1/CHIP contributes to the Th17/Treg imbalance via the ubiquitination of aryl hydrocarbon receptor in rheumatoid arthritis

Wang, Wen; Xiang, Ting; Yang, Yachen; Wang, Zitao; Xie, Jianmin

doi:10.1093/cei/uxac072

Cited by 16 publications

(8 citation statements)

References 42 publications

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“…GO annotation analysis showed ubiquitin mediated proteolysis as the most significantly enriched pathway (p<0.0001) in presence of MG132 (Supplementary figure 6A-C). Previously known substrates such as SQSTM1( 65 ), NPM1( 67 ), PSMD4( 68 ) and HDAC6( 69 ) were identified in this Ub-POD analysis (Supplementary table 8). Applying Ub-POD to CHIP demonstrated that a flexible linker between BirA and the candidate Ub ligase can improve the identification of substrates and that biotin exposure times and stimuli may need to be optimized for a given Ub ligase to identify different sets of substrates.…”

Section: Resultsmentioning

confidence: 90%

“…Therefore, instead of exposing the cells to biotin immediately after transfection, we added biotin to cells 24h after the transfection, along with MG132 (Supplementary Figure 3A). Interestingly, two other known substrates of CHIP, SQSTM1 (also known as p62) (57) and SOD2 (58) were identified (Supplementary Figure 3B, Supplementary table 8) with this short biotin exposure. This change in substrate identification profile depending on the duration of biotin pulse could be due to the different half-lives of various substrates of the CHIP Ub ligase.…”

Section: Employing Ub-pod To Identify Substrates Of U-box Domain Cont...mentioning

confidence: 97%

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A ubiquitin-specific, proximity-based labeling approach for the identification of ubiquitin ligase substrates

Mukhopadyay,

Levantovsky,

Gharbi

et al. 2023

Preprint

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Ubiquitination of proteins is central to protein homeostasis and other cellular processes including DNA repair, vesicular transport, cell-division etc. The process of ubiquitination is conserved from yeast to humans and is carried out by the sequential action of three enzymes: E1, E2 and E3. There are an estimated >600 E3 ligases in humans that execute ubiquitination of specific target proteins in a spatio-temporal manner to elicit desired signaling effects. Here, we developed a simple, proximity-based labeling method to selectively biotinylate substrates of a given ubiquitin ligase. Our method exploits the proximity and the relative orientation of the E3-ligase catalytic domain with respect to ubiquitin observed in the enzymatic intermediate-state structures of E3-E2~Ub. By fusing the biotin ligase BirA and an Avi tag variant to the candidate E3 ligase and ubiquitin, respectively, we were able to specifically enrich the bona fide substrates and potential new substrates of the ligase using a one-step Streptavidin pulldown under denaturing conditions. As proof-of-principle, we applied our method, which we named Ub-POD, to the RING E3 ligase Rad18. Rad18 ubiquitinates DNA-sliding clamp PCNA upon UV-induced DNA damage. We identified PCNA and several other critical players in the DNA damage repair pathway in a single Ub-POD experiment. We further discovered an unintended but important application of Ub-POD, where we were able to pin down the cellular localization of Rad18-mediated ubiquitination to the damaged DNA nuclear puncta through Streptavidin immunofluorescence. We also applied our method to TRAF6, another RING ubiquitin ligase involved in TNF signaling, successfully identifying known and potentially new substrates. Finally, we adapted our method to the U-box-type E3 ubiquitin ligase CHIP to demonstrate that we can identify substrates of two major classes of mammalian ubiquitin ligases. We anticipate that our method and principle could be widely adapted to all classes of ubiquitin ligases to identify substrates and localize the cellular site(s) of ubiquitination.

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Section: Resultsmentioning

confidence: 90%

Section: Employing Ub-pod To Identify Substrates Of U-box Domain Cont...mentioning

confidence: 97%

A ubiquitin-specific, proximity-based labeling approach for the identification of ubiquitin ligase substrates

Mukhopadyay,

Levantovsky,

Gharbi

et al. 2023

Preprint

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“…AHR plays a key regulatory role in the differentiation of Treg and Th17 cells. Studies have found that the expression of AhR in CD4 + T cells leads to a signi cant increase in the percentage of Treg and related gene transcripts (including Foxp3, IL-10, and CD39), and a signi cant decrease in the transcription of Th17 cells and related genes (including RORγt, IL-17A, and IL-17F), inducing an imbalance between Th17 and Treg cells [49,50].…”

Section: Discussionmentioning

confidence: 99%

The effects of thioredoxin peroxidase from Cysticercus cellulosae excretory-secretory antigens on TGF-β signaling pathway and Th17 cells differentiation in Jurkat cells by transcriptomics

Sun

Yang

et al. 2023

Preprint

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(1) Background: Thioredoxin peroxidase (TPx) protein from the excretory-secretory antigens (ESAs) of Cysticercus cellulosae (C. cellulosae) has been shown to regulate the differentiation of host Treg and Th17 cells, resulting in an immunosuppressive response dominated by Treg cells. However, the molecular mechanism by which TPx protein from the ESAs of C. cellulosae regulates the imbalance of host Treg/Th17 cell differentiation has not been reported. (2) Methods: TPx protein from porcine C. cellulosae ESAs was used to stimulate Jurkat cells activated with PMA and Ionomycin at 0, 24, 48, and 72 hours. Transcriptomic analysis was performed to investigate the signaling pathways associated with Jurkat cells differentiation regulated by TPx protein from C. cellulosae ESAs. (3) Results: Gene Set Enrichment Analysis (GSEA) revealed that TPx protein from porcine C. cellulosae ESAs could induce upregulation of the TGF-β signaling pathway and downregulation of Th17 cell differentiation in Jurkat cells. (4) Conclusion: TPx protein from porcine C. cellulosae ESAs can activate the TGF-β signaling pathway in Jurkat cells, thereby regulating the differentiation of Treg/Th17 cells and leading to an immunosuppressive response dominated by Treg cells, enabling evasion of the host immune attack. This study provides a foundation for further validation of these pathways and further elucidates the molecular mechanisms underlying immune evasion caused by porcine C. cellulosae.

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“…The new RA pathogenesis hypothesis suggests that Th17/Treg and Th1/Th2 imbalance may be the cause of RA (30) . Initially, Th1 cells have a pathogenic effect on RA.…”

Section: Discussionmentioning

confidence: 99%

hIgD-Fc-Ig fusion protein regulate T cell functions by inhibiting TCR signaling pathway in adjuvant arthritis rats

Zhang

Wang

et al. 2022

Preprint

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hIgD-Fc-Ig is a fusion protein that competes to bind IgD receptors, it remains unclear whether hIgD-Fc-Ig can regulate T cell function by regulating TCR signaling pathway in the treatment of adjuvant arthritis rats. In vivo, AA rats were treated with hIgD-Fc-Ig fusion protein and Etanercept for 28 days, then the overall indexes of AA rats, the severity of the pathology, the proliferation of spleen and thymus, the changes of blood flow signal in the knee joints as well as bone erosion of ankle joints were detected. Flow cytometry was used to detect the changes of peripheral blood and spleen T cell subsets. In vitro, rat spleen T cells or Jurkat cells were treated by IgD, and Lck inhibitor (PP1) and CD3ε siRNA were used to observe the function of IgD and hIgD-Fc-Ig on TCR and its downstream protein expression. The results showed that hIgD-Fc-Ig fusion protein had a obvious therputic effect on adjuvant arthritis rats, which could improve overall index, pathological status, the proportion of T cell subsets and other indicators. In addition, hIgD-Fc-Ig inhibited the expression of TCR and its downstream related proteins in rat spleen T cells or Jurkat cells. Which provided evidence that hIgD-Fc-Ig fusion protein could alleviate the symptoms of AA rats and regulate T cells through TCR-Lck-Erk signaling pathway. In a word, activated TCR signaling pathway leads to T cell activation which could be inhibited by hIgD-Fc-Ig fusion protein through regulating TCR signaling pathway. hIgD-Fc-Ig might be an immunomodulatory drug with anti-inflammatory effects.

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E3 ubiquitin ligases STUB1/CHIP contributes to the Th17/Treg imbalance via the ubiquitination of aryl hydrocarbon receptor in rheumatoid arthritis

Cited by 16 publications

References 42 publications

A ubiquitin-specific, proximity-based labeling approach for the identification of ubiquitin ligase substrates

A ubiquitin-specific, proximity-based labeling approach for the identification of ubiquitin ligase substrates

The effects of thioredoxin peroxidase from Cysticercus cellulosae excretory-secretory antigens on TGF-β signaling pathway and Th17 cells differentiation in Jurkat cells by transcriptomics

hIgD-Fc-Ig fusion protein regulate T cell functions by inhibiting TCR signaling pathway in adjuvant arthritis rats

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