The cytokine TNF can trigger highly proinflammatory RIPK1-dependent cell death. Here, we show that the two adapter proteins, TANK and AZI2, suppress TNF-induced cell death by regulating the activation of TBK1 kinase. Mice lacking either TANK or AZI2 do not show an overt phenotype. Conversely, animals deficient in both adapters are born in a sub-Mendelian ratio and suffer from severe multi-organ inflammation, excessive antibody production, male sterility, and early mortality, which can be rescued by TNFR1 deficiency and significantly improved by expressing a kinase-dead form of RIPK1. Mechanistically, TANK and AZI2 both recruit TBK1 to the TNF receptor signaling complex, but with distinct kinetics due to interaction with different complex components. While TANK binds directly to the adapter NEMO, AZI2 is recruited later via deubiquitinase A20. In summary, our data show that TANK and AZI2 cooperatively sustain TBK1 activity during different stages of TNF receptor assembly to protect against autoinflammation.
- MeSH
- adaptorové proteiny signální transdukční * metabolismus genetika MeSH
- buněčná smrt MeSH
- endopeptidasy MeSH
- intracelulární signální peptidy a proteiny metabolismus genetika MeSH
- lidé MeSH
- myši inbrední C57BL MeSH
- myši knockoutované * MeSH
- myši MeSH
- protein-serin-threoninkinasy * metabolismus genetika MeSH
- receptory TNF - typ I * metabolismus genetika MeSH
- serin-threoninkinasy interagující s receptory * metabolismus genetika MeSH
- signální transdukce MeSH
- TNF-alfa * metabolismus MeSH
- TNFAIP3 metabolismus genetika MeSH
- zánět metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Radiation-induced intestinal injury (RIII) interrupts the scheduled processes of abdominal and pelvic radiotherapy (RT) and compromises the quality of life of cancer survivors. However, the specific regulators and mechanisms underlying the effects of RIII remain unknown. The biological effects of RT are caused primarily by DNA damage, and ataxia telangiectasia mutated (ATM) is a core protein of the DNA damage response (DDR). However, whether ATM is regulated by deubiquitination signaling remains unclear. METHODS: We established animal and cellular models of RIII. The effects of ubiquitin-specific protease 15 (USP15) on DNA damage and radion-induced intestinal injury were evaluated. Mass spectrometry analysis, truncation tests, and immunoprecipitation were used to identify USP15 as a binding partner of ATM and to investigate the ubiquitination of ATM. Finally, the relationship between the USP15/ATM axes was further determined via subsequent experiments. RESULTS: In this study, we identified the deubiquitylating enzyme USP15 as a regulator of DNA damage and the pathological progression of RIII. Irradiation upregulates the expression of USP15, whereas pharmacological inhibition of USP15 exacerbates radiation-induced DNA damage and RIII both in vivo and in vitro. Mechanistically, USP15 interacts with, deubiquitinates, and stabilises ATM via K48-linked deubiquitination. Notably, ATM overexpression blocks the effect of USP15 genetic inhibition on DNA damage and RIII progression. CONCLUSIONS: These findings describe ATM as a novel deubiquitination target of USP15 upon radiation-induced DNA damage and intestinal injury, and provides experimental support for USP15/ATM axis as a potential target for developing strategies that mitigate RIII.
- MeSH
- ATM protein * metabolismus genetika MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- poškození DNA * MeSH
- radiační poranění metabolismus genetika MeSH
- specifické proteázy ubikvitinu * metabolismus genetika MeSH
- střeva účinky záření patologie MeSH
- ubikvitinace * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Serum monoclonal immunoglobulin (Ig) is the main diagnostic factor for patients with multiple myeloma (MM), however its prognostic potential remains unclear. On a large MM patient cohort (n = 4146), we observe no correlation between serum Ig levels and patient survival, while amount of intracellular Ig has a strong predictive effect. Focused CRISPR screen, transcriptional and proteomic analysis identify deubiquitinase OTUD1 as a critical mediator of Ig synthesis, proteasome inhibitor sensitivity and tumor burden in MM. Mechanistically, OTUD1 deubiquitinates peroxiredoxin 4 (PRDX4), protecting it from endoplasmic reticulum (ER)-associated degradation. In turn, PRDX4 facilitates Ig production which coincides with the accumulation of unfolded proteins and higher ER stress. The elevated load on proteasome ultimately potentiates myeloma response to proteasome inhibitors providing a window for a rational therapy. Collectively, our findings support the significance of the Ig production machinery as a biomarker and target in the combinatory treatment of MM patients.
- MeSH
- apoptóza MeSH
- bortezomib farmakologie terapeutické užití MeSH
- deubikvitinasy MeSH
- imunoglobuliny MeSH
- inhibitory proteasomu * farmakologie MeSH
- lidé MeSH
- mnohočetný myelom * farmakoterapie genetika metabolismus MeSH
- proteasomový endopeptidasový komplex metabolismus MeSH
- proteomika MeSH
- specifické proteázy ubikvitinu MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Genetic defects in the repair of DNA single-strand breaks (SSBs) can result in neurological disease triggered by toxic activity of the single-strand-break sensor protein PARP1. However, the mechanism(s) by which this toxic PARP1 activity triggers cellular dysfunction are unclear. Here we show that human cells lacking XRCC1 fail to rapidly recover transcription following DNA base damage, a phenotype also observed in patient-derived fibroblasts with XRCC1 mutations and Xrcc1-/- mouse neurons. This defect is caused by excessive/aberrant PARP1 activity during DNA base excision repair, resulting from the loss of PARP1 regulation by XRCC1. We show that aberrant PARP1 activity suppresses transcriptional recovery during base excision repair by promoting excessive recruitment and activity of the ubiquitin protease USP3, which as a result reduces the level of monoubiquitinated histones important for normal transcriptional regulation. Importantly, inhibition and/or deletion of PARP1 or USP3 restores transcriptional recovery in XRCC1-/- cells, highlighting PARP1 and USP3 as possible therapeutic targets in neurological disease.
- MeSH
- DNA genetika MeSH
- genetická transkripce genetika MeSH
- histony metabolismus MeSH
- jednořetězcové zlomy DNA * MeSH
- lidé MeSH
- myši knockoutované MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- oprava DNA genetika MeSH
- oxidační stres genetika MeSH
- peroxid vodíku toxicita MeSH
- poly(ADP-ribosa)polymerasa 1 genetika metabolismus MeSH
- protein XRCC1 genetika metabolismus MeSH
- specifické proteázy ubikvitinu metabolismus MeSH
- ubikvitinace fyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A common strategy for exploring the biological roles of deubiquitinating enzymes (DUBs) in different pathways is to study the effects of replacing the wild-type DUB with a catalytically inactive mutant in cells. We report here that a commonly studied DUB mutation, in which the catalytic cysteine is replaced with alanine, can dramatically increase the affinity of some DUBs for ubiquitin. Overexpression of these tight-binding mutants thus has the potential to sequester cellular pools of monoubiquitin and ubiquitin chains. As a result, cells expressing these mutants may display unpredictable dominant negative physiological effects that are not related to loss of DUB activity. The structure of the SAGA DUB module bound to free ubiquitin reveals the structural basis for the 30-fold higher affinity of Ubp8C146A for ubiquitin. We show that an alternative option, substituting the active site cysteine with arginine, can inactivate DUBs while also decreasing the affinity for ubiquitin.
- MeSH
- alanin genetika MeSH
- cystein genetika MeSH
- deubikvitinasy chemie genetika MeSH
- endopeptidasy chemie genetika MeSH
- katalýza MeSH
- konformace proteinů MeSH
- lidé MeSH
- mutace genetika MeSH
- Saccharomyces cerevisiae - proteiny chemie genetika MeSH
- Saccharomyces cerevisiae genetika MeSH
- specifické proteázy ubikvitinu chemie genetika MeSH
- substituce aminokyselin genetika MeSH
- trans-aktivátory chemie genetika MeSH
- transportní proteiny chemie genetika MeSH
- ubikvitin chemie genetika MeSH
- ubikvitinace genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
The overexpression of Mdm2 has been linked to the loss of p53 tumour suppressor activity in several human cancers. Here, we present results suggesting that ubiquitin-specific peptidase 48 (USP48), a deubiquitinase that has been linked in previous reports to the NF-κB signaling pathway, is a novel Mdm2 binding partner that promotes Mdm2 stability and enhances Mdm2-mediated p53 ubiquitination and degradation. In contrast to other deubiquitinating enzymes (DUBs) that have been previously implicated in the regulation of Mdm2 protein stability, USP48 did not induce Mdm2 stabilization by significantly reducing Mdm2 ubiquitination levels. Moreover, two previously characterized USP48 mutants lacking deubiquitinase activity were also capable of efficiently stabilizing Mdm2, indicating that USP48 utilizes a non-canonical, deubiquitination-independent mechanism to promote Mdm2 oncoprotein stability. This study represents, to the best of our knowledge, the first report suggesting DUB-mediated target protein stabilization that is independent of its deubiquitinase activity. In addition, our results suggest that USP48 might represent a new mechanism of crosstalk between the NF-κB and p53 stress response pathways.
- MeSH
- buněčné linie MeSH
- lidé MeSH
- nádorový supresorový protein p53 metabolismus MeSH
- posttranslační úpravy proteinů MeSH
- proteolýza MeSH
- protoonkogenní proteiny c-mdm2 metabolismus MeSH
- regulace genové exprese * MeSH
- specifické proteázy ubikvitinu metabolismus MeSH
- ubikvitinace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- dexamethason aplikace a dávkování MeSH
- lidé MeSH
- mnohočetný myelom * farmakoterapie MeSH
- náklady na léky MeSH
- přežití bez známek nemoci MeSH
- protokoly protinádorové kombinované chemoterapie MeSH
- specifické proteázy ubikvitinu antagonisté a inhibitory aplikace a dávkování MeSH
- thalidomid analogy a deriváty aplikace a dávkování MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- klinické zkoušky MeSH
- novinové články MeSH
INTRODUCTION: Deubiquitinating-enzymes (DUBs) are key components of the ubiquitin-proteasome system (UPS). The fundamental role of DUBs is specific removal of ubiquitin from substrates. DUBs contribute to activation/deactivation, recycling and localization of numerous regulatory proteins, and thus play major roles in diverse cellular processes. Altered DUB activity is associated with a multitudes of pathologies including cancer. Therefore, DUBs represent novel candidates for target-directed drug development. AREAS COVERED: The article is a thorough review/accounting of patented compounds targeting DUBs and stratifying/classifying the patented compounds based on: chemical-structures, nucleic-acid compositions, modes-of-action, and targeting sites. The review provides a brief background on the UPS and the involvement of DUBs. Furthermore, methods for assessing efficacy and potential pharmacological utility of DUB inhibitor (DUBi) are discussed. EXPERT OPINION: The FDA's approval of the 20S proteasome inhibitors (PIs): bortezomib and carfilzomib for treatment of hematological malignancies established the UPS as an anti-cancer target. Unfortunately, many patients are inherently resistant or develop resistance to PIs. One potential strategy to combat PI resistance is targeting upstream components of the UPS such as DUBs. DUBs represent a promising potential therapeutic target due to their critical roles in various cellular processes including protein turnover, localization and cellular homeostasis. While considerable efforts have been undertaken to develop DUB modulators, significant advancements are necessary to move DUBis into the clinic.
- MeSH
- chemorezistence MeSH
- cílená molekulární terapie MeSH
- inhibitory proteasomu farmakologie MeSH
- lidé MeSH
- nádory farmakoterapie enzymologie MeSH
- patenty jako téma MeSH
- protinádorové látky farmakologie MeSH
- racionální návrh léčiv MeSH
- specifické proteázy ubikvitinu antagonisté a inhibitory metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Research Support, N.I.H., Extramural MeSH
Huntingtonova nemoc je progresivní dědičné neurodegenerativní onemocnění s autozomálně dominantním typem dědičnosti a nástupem klinických příznaků okolo 30 – 50 roku života. Podkladem nemoci je mutace genu pro protein huntingtin. Dochází ke zmnožení CAG repetic a syntéze polyglutaminu. Vlivem odumírání neuronů se objevuje mozková atrofie, pacient trpí změnami nálad, zhoršením kognitivních a motorických funkcí, depresemi. Nakonec dochází k celkovému rozpadu osobnosti.
Huntington‘s disease is a hereditary progressive neurodegenerative disease with an autosomal dominant pattern of inheritance and the onset of clinical symptoms about 30 to 50 years of age. A basis of the disease is a mutation of gene for a huntingtin protein. There is a multiplication of CAG repeats and the synthesis of polyglutamine. Due to death of neurons, a cerebral atrophy occurs, patients suffer from mood swings, impairment of cognitive and motor functions, depression. Finally, there is an overall personality breakdown.
- MeSH
- amyloidogenní proteiny metabolismus škodlivé účinky MeSH
- apoptóza fyziologie MeSH
- centrální nervový systém enzymologie patofyziologie patologie MeSH
- genetické pozadí MeSH
- lidé MeSH
- neurodegenerativní nemoci etiologie klasifikace patofyziologie MeSH
- neurony * fyziologie patologie MeSH
- proteinové agregáty * fyziologie genetika imunologie MeSH
- reaktivní formy kyslíku škodlivé účinky MeSH
- specifické proteázy ubikvitinu metabolismus škodlivé účinky MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
