Structure of the Rpn13-Rpn2 complex provides insights for Rpn13 and Uch37 as anticancer targets

Lu, Xiuxiu; Nowicka, Urszula; Sridharan, Vinidhra; Liu, Fen; Randles, Leah; Hymel, David; Dyba, Marcin; Tarasov, Sergey G.; Tarasova, Nadya I.; Zhao, Xue Zhi; Hamazaki, Jun; Murata, Shigeo; Burke, Terrence R.; Walters, Kylie J.

doi:10.1038/ncomms15540

Cited by 69 publications

(144 citation statements)

References 70 publications

(133 reference statements)

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“…RA190, which can covalently bind to cysteine 88 of ADRM1 in the 19S regulatory particle of proteasome, inhibits proteasome function, resulting in higher-molecular weight polyubiquitylated protein accumulation [14,19]. In HCT116 cells, RA190 can also bind with UCH37, a deubiquitinating enzyme interacting with ADRM1, to suppress proteasome function [35]. Meanwhile, it was reported that KDT-11 could also suppress multiple myeloma by targeting ADRM1 [36].…”

Section: Discussionmentioning

confidence: 99%

RA190, a Proteasome Subunit ADRM1 Inhibitor, Suppresses Intrahepatic Cholangiocarcinoma by Inducing NF-KB-Mediated Cell Apoptosis

Wang

Zheng

et al. 2018

Cell Physiol Biochem

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Background/Aims: Effective drug treatment for intrahepatic cholangiocarcinoma (ICC) is currently lacking. Therefore, there is an urgent need for new targets and new drugs that can prolong patient survival. Recently targeting the ubiquitin proteasome pathway has become an attractive anti-cancer strategy. In this study, we aimed to evaluate the therapeutic effect of and identify the potential mechanisms involved in targeting the proteasome subunit ADRM1 for ICC. Methods: The expression of ADRM1 and its prognostic value in ICC was analyzed using GEO and TCGA datasets, tumor tissues, and tumor tissue arrays. The effects of RA190 on the proliferation and survival of both established ICC cell lines and primary ICC cells were examined in vitro. Annexin V/propidium iodide staining, western blotting and immunohistochemical staining were performed. The in vivo anti-tumor effect of RA190 on ICC was validated in subcutaneous xenograft and patient-derived xenograft (PDX) models. Results: ADRM1 levels were significantly higher in ICC tissues than in normal bile duct tissues. ICC patients with high ADRM1 levels had worse overall survival (hazard ratio [HR] = 2.383, 95% confidence interval [CI] =1.357 to 4.188) and recurrence-free survival (HR = 1.710, 95% CI =1.045 to 2.796). ADRM1 knockdown significantly inhibited ICC growth in vitro and in vivo. The specific inhibitor RA190 targeting ADRM1 suppressed proliferation and reduced cell vitality of ICC cell lines and primary ICC cells significantly in vitro. Furthermore, RA190 significantly inhibited the proteasome by inactivating ADRM1, and the consequent accumulation of ADRM1 substrates decreased the activating levels of NF-κB to aggravate cell apoptosis. The therapeutic benefits of RA190 treatment were further demonstrated in both subcutaneous implantation and PDX models. Conclusions: Our findings indicate that up-regulated ADRM1 was involved in ICC progression and suggest the potential clinical application of ADRM1 inhibitors (e.g., RA190 and KDT-11) for ICC treatment.

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Section: Discussionmentioning

confidence: 99%

RA190, a Proteasome Subunit ADRM1 Inhibitor, Suppresses Intrahepatic Cholangiocarcinoma by Inducing NF-KB-Mediated Cell Apoptosis

Wang

Zheng

et al. 2018

Cell Physiol Biochem

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“…RP-associated Uch37 was found to progressively trim ubiquitin chains from the distal end, removing one ubiquitin moiety at a time [96,97]. Both proteasome-associated Uch37 and the isolated Uch37 catalytic domain have been shown to act on chains of varied linkage types [88,98,99], and RP-associated Uch37 has been reported to remove ubiquitin moieties from α-globin [96]. The efficiency of Uch37-mediated hydrolysis, however, seems strikingly low: whereas Usp14 acts on its preferred substrates within seconds [35], Uch37-mediated cleavage of Ub-chains takes much longer, with incubations of 30 minutes and even 8 hours [88,96,98,99].…”

Section: Substrate Specificitymentioning

confidence: 99%

“…Both proteasome-associated Uch37 and the isolated Uch37 catalytic domain have been shown to act on chains of varied linkage types [88,98,99], and RP-associated Uch37 has been reported to remove ubiquitin moieties from α-globin [96]. The efficiency of Uch37-mediated hydrolysis, however, seems strikingly low: whereas Usp14 acts on its preferred substrates within seconds [35], Uch37-mediated cleavage of Ub-chains takes much longer, with incubations of 30 minutes and even 8 hours [88,96,98,99]. It may be suggested that Uch37 acts slowly in order to provide the proteasome with ample time to degrade bona fide substrates [60], slowly deubiquitinating substrates that stall the proteasome until they are released.…”

Section: Substrate Specificitymentioning

confidence: 99%

Meddling with Fate: The Proteasomal Deubiquitinating Enzymes

Poot

Tian

Finley

2017

Journal of Molecular Biology

103

115

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Three deubiquitinating enzymes–Rpn11, Usp14, and Uch37–are associated with the proteasome regulatory particle. These enzymes allow proteasomes to remove ubiquitin from substrates before they are translocated into the core particle to be degraded. Although the translocation channel is too narrow for folded proteins, the force of translocation unfolds them mechanically. As translocation proceeds, ubiquitin chains bound to substrate are drawn to the channel’s entry port, where they can impede further translocation. Rpn11, situated over the port, can remove these chains without compromising degradation because substrates must be irreversibly committed to degradation before Rpn11 acts. This coupling between deubiquitination and substrate degradation is ensured by the Ins-1 loop of Rpn11, which controls ubiquitin access to its catalytic site. In contrast to Rpn11, Usp14 and Uch37 can rescue substrates from degradation by promoting substrate dissociation from the proteasome prior to the commitment step. Uch37 is unique in being a component of both the proteasome and a second multisubunit assembly, the INO80 complex. However, only recruitment into the proteasome activates Uch37. Recruitment to the proteasome likewise activates Usp14. However, the influence of Usp14 on the proteasome depends on the substrate, due to its marked preference for proteins that carry multiple ubiquitin chains. Usp14 exerts complex control over the proteasome, suppressing proteasome activity even when inactive in deubiquitination. A major challenge for the field will be to elucidate the specificities of Rpn11, Usp14, and Uch37 in greater depth, employing not only model in vitro substrates, but their endogenous targets.

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“…How might binding of polyubiquitin be communicated from the receptors to the Rpt motor proteins? Rpn13 is connected to the proteasome via attachment to a flexible extension at the C-terminus of Rpn2 (Figure 8A), and is fairly poorly resolved in cryoEM structures, suggesting that a range of conformations may be possible 58 . Ubiquitin occupancy may then be communicated to the ring of Rpt motor proteins via these different conformations.…”

Section: Discussionmentioning

confidence: 99%

Ubiquitin receptors are required for substrate-mediated activation of the proteasome’s unfolding ability

Cundiff

Hurley

Wong

et al. 2019

Preprint

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The ubiquitin-proteasome system (UPS) is responsible for the bulk of protein degradation in eukaryotic cells, but the factors that cause different substrates to be unfolded and degraded to different extents are still poorly understood. We previously showed that polyubiquitinated substrates were degraded with greater processivity (with a higher tendency to be unfolded and degraded than released) than ubiquitin-independent substrates. Thus, even though ubiquitin chains are removed before unfolding and degradation occur, they affect the unfolding of a protein domain. How do ubiquitin chains activate the proteasome's unfolding ability? We investigated the roles of the three intrinsic proteasomal ubiquitin receptors -Rpn1, Rpn10 and Rpn13 -in this activation. We find that these receptors are required for substrate-mediated activation of the proteasome's unfolding ability. Rpn13 plays the largest role, but there is also partial redundancy between receptors. The architecture of substrate ubiquitination determines which receptors are needed for maximal unfolding ability, and, in some cases, simultaneous engagement of ubiquitin by multiple receptors may be required. Our results suggest physical models for how ubiquitin receptors communicate with the proteasomal motor proteins.

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Structure of the Rpn13-Rpn2 complex provides insights for Rpn13 and Uch37 as anticancer targets

Cited by 69 publications

References 70 publications

RA190, a Proteasome Subunit ADRM1 Inhibitor, Suppresses Intrahepatic Cholangiocarcinoma by Inducing NF-KB-Mediated Cell Apoptosis

RA190, a Proteasome Subunit ADRM1 Inhibitor, Suppresses Intrahepatic Cholangiocarcinoma by Inducing NF-KB-Mediated Cell Apoptosis

Meddling with Fate: The Proteasomal Deubiquitinating Enzymes

Ubiquitin receptors are required for substrate-mediated activation of the proteasome’s unfolding ability

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