The Regulations of Deubiquitinase USP15 and Its Pathophysiological Mechanisms in Diseases

Chou, Chia-Man; Chang, Yu-Ting; Korinek, Michal; Chen, Yei Tsung; Yang, Yating; Leu, Steve; Lin, I‐Ling; Tang, Chin-Ju; Chiu, Chien‐Chih

doi:10.3390/ijms18030483

Cited by 44 publications

(33 citation statements)

References 80 publications

(116 reference statements)

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“…USP15 belongs to the Ub-specific protease (USP) family, the largest DUB structural family containing $56 members in humans, and shares the same domain architecture and high sequence similarity with two homologs, USP4 (57% similarity) and USP11 (43% similarity) (Chou et al, 2017). In addition to the catalytic domain, USP15 contains a DUSP domain (domain present in Ub-specific proteases), which has no described function and is found exclusively in USP proteins (Clague et al, 2013), and two Ub-like (Ubl) domains, which share the conserved b-grasp fold of Ub (Burroughs et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15

et al. 2019

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Highlights d Tight and selective UbVs target USP15 catalytic and adaptor domains d UbV inhibitors lock the USP15 active site in an inactive conformation d A strand-swapped UbV dimer binds two DUSP domains simultaneously d Linear UbV dimers are potent and specific USP15 inhibitors in cells SUMMARYThe multi-domain deubiquitinase USP15 regulates diverse eukaryotic processes and has been implicated in numerous diseases. We developed ubiquitin variants (UbVs) that targeted either the catalytic domain or each of three adaptor domains in USP15, including the N-terminal DUSP domain. We also designed a linear dimer (diUbV), which targeted the DUSP and catalytic domains, and exhibited enhanced specificity and more potent inhibition of catalytic activity than either UbV alone. In cells, the UbVs inhibited the deubiquitination of two USP15 substrates, SMURF2 and TRIM25, and the diUbV inhibited the effects of USP15 on the transforming growth factor b pathway. Structural analyses revealed that three distinct UbVs bound to the catalytic domain and locked the active site in a closed, inactive conformation, and one UbV formed an unusual strand-swapped dimer and bound two DUSP domains simultaneously. These inhibitors will enable the study of USP15 function in oncology, neurology, immunology, and inflammation.

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Section: Introductionmentioning

confidence: 99%

Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15

et al. 2019

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“…Structural information is key to shedding light on their mechanism and substrate interactions and can aid small-molecule inhibitor development. The multifunctional protease USP15 regulates several important pathways in health and disease ( 4 ), including TGF-β ( 5 ), IGF-I ( 6 ), innate immune signaling ( 7 ), mRNA processing ( 8 ), and mitophagy ( 9 ). Furthermore, USP15 can promote new protein synthesis ( 10 ), depletion results in DNA double strand repair defects ( 11 ), and USP15 was shown to regulate the ligase MDM2 with effects on the stability of p53 in cancer cells and the T-cell transcription factor, NFATc2 ( 12 ).…”

Section: Introductionmentioning

confidence: 99%

The structure of the deubiquitinase USP15 reveals a misaligned catalytic triad and an open ubiquitin-binding channel

Ward

Gratton

Indrayudha

et al. 2018

Journal of Biological Chemistry

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Ubiquitin-specific protease 15 (USP15) regulates important cellular processes, including transforming growth factor β (TGF-β) signaling, mitophagy, mRNA processing, and innate immune responses; however, structural information on USP15's catalytic domain is currently unavailable. Here, we determined crystal structures of the USP15 catalytic core domain, revealing a canonical USP fold, including a finger, palm, and thumb region. Unlike for the structure of paralog USP4, the catalytic triad is in an inactive configuration with the catalytic cysteine ∼10 Å apart from the catalytic histidine. This conformation is atypical, and a similar misaligned catalytic triad has so far been observed only for USP7, although USP15 and USP7 are differently regulated. Moreover, we found that the active-site loops are flexible, resulting in a largely open ubiquitin tail–binding channel. Comparison of the USP15 and USP4 structures points to a possible activation mechanism. Sequence differences between these two USPs mainly map to the S1′ region likely to confer specificity, whereas the S1 ubiquitin–binding pocket is highly conserved. Isothermal titration calorimetry monoubiquitin- and linear diubiquitin-binding experiments showed significant differences in their thermodynamic profiles, with USP15 displaying a lower affinity for monoubiquitin than USP4. Moreover, we report that USP15 is weakly inhibited by the antineoplastic agent mitoxantrone in vitro. A USP15–mitoxantrone complex structure disclosed that the anthracenedione interacts with the S1′ binding site. Our results reveal first insights into USP15's catalytic domain structure, conformational changes, differences between paralogs, and small-molecule interactions and establish a framework for cellular probe and inhibitor development.

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“…USP15 is previously reported to be dysregulated in many human cancers and plays critical roles in tumor development and progression [17]. Here, we first analyzed USP15 gene expression in different types of human leukemia using The Cancer Genome Atlas (TCGA) database.…”

Section: Usp15 Expression Is Significantly Downregulated In CMLmentioning

confidence: 99%

De-regulated STAT5A/miR-202-5p/USP15/Caspase-6 regulatory axis suppresses CML cell apoptosis and contributes to Imatinib resistance

Nie

Yao

Yang

et al. 2020

J Exp Clin Cancer Res

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Background: STAT5 plays an important role in the transformation of hematopoietic cells by BCR-ABL. However, the downstream target genes activated by STAT5 in chronic myeloid leukemia (CML) cells remain largely unclear. Here, we investigated the mechanistic functional relationship between STAT5A-regulated microRNA and CML cell apoptosis. Methods: The expression of USP15, Caspase-6, STAT5A-regulated miR-202-5p and STAT5A was detected by qRT-PCR and Western blotting in CML cell lines and PBMCs of CML patients. Cell apoptosis was evaluated by flow cytometry. Both gain-and loss-of-function experiments were used to investigate the roles of USP15, miR-202-5p and STAT5A in CML. Luciferase reporter assay detected the effect of miR-202-5p on USP15 expression. Xenograft animal model was used to test the effect of anti-miR-202-5p and pimozide on K562 cell xenograft growth. Results: USP15 expression was significantly downregulated in CML cell lines and PBMCs of CML patients. Depletion of USP15 increased, whereas overexpression of USP15 reduced the resistance of CML cells to Imatinib. Further, decreased deubiquitinating activity of USP15 by USP15 downregulation led to reduced caspase-6 level, thus attenuating CML cell apoptosis. Mechanistically, miR-202-5p was upregulated in K562G cells and negatively regulated USP15 expression by directly targeting USP15 3′-UTR. Correspondingly, upregulation of miR-202-5p enhanced the resistance of CML cells to Imatinib by inhibiting cell apoptosis. Importantly, STAT5A was upregulated in CML cells and directly activated miR-202-5p transcription by binding to the pre-miR-202 promoter. Pimozide induced CML cell apoptosis and significantly reduced K562 cell xenograft growth in vivo by blocking STAT5A/miR-202-5p/USP15/Caspase-6 regulatory axis. Conclusions: we provide the first evidence that de-regulated STAT5A/miR-202-5p/USP15/Caspase-6 regulatory axis suppresses the apoptosis of CML cells, targeting this pathway might be a promising therapeutic approach for the treatment of CML.

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The Regulations of Deubiquitinase USP15 and Its Pathophysiological Mechanisms in Diseases

Cited by 44 publications

References 80 publications

Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15

Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15

The structure of the deubiquitinase USP15 reveals a misaligned catalytic triad and an open ubiquitin-binding channel

De-regulated STAT5A/miR-202-5p/USP15/Caspase-6 regulatory axis suppresses CML cell apoptosis and contributes to Imatinib resistance

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