RESUMEN

Red-spotted grouper nervous necrosis virus (RGNNV) is a major viral pathogen of grouper and is able to antagonize interferon responses through multiple strategies, particularly evading host immune responses by inhibiting interferon responses. Ovarian tumor (OTU) family proteins are an important class of DUBs and the underlying mechanisms used to inhibit interferon pathway activation are unknown. In the present study, primers were designed based on the transcriptome data, and the ovarian tumor (OTU) domain-containing ubiquitin aldehyde-binding protein 1 (OTUB1) and OTUB2 genes of Epinephelus coioides (EcOTUB1 and EcOTUB2) were cloned and characterized. The homology alignment showed that both EcOTUB1 and EcOTUB2 were most closely related to E. lanceolatus with 98 % identity. Both EcOTUB1 and EcOTUB2 were distributed to varying degrees in grouper tissues, and the transcript levels were significantly up-regulated following RGNNV stimulation. Both EcOTUB1 and EcOTUB2 promoted replication of RGNNV in vitro, and inhibited the promoter activities of interferon stimulated response element (ISRE), nuclear transcription factors kappaB (NF-κB) and IFN3, and the expression levels of interferon related genes and proinflammatory factors. Co-immunoprecipitation experiments showed that both EcOTUB1 and EcOTUB2 could interact with TRAF3 and TRAF6, indicating that EcOTUB1 and EcOTUB2 may play important roles in interferon signaling pathway. The results will provide a theoretical reference for the development of novel disease prevention and control techniques.

Asunto(s)

Lubina , Enfermedades de los Peces , Proteínas de Peces , Inmunidad Innata , Nodaviridae , Infecciones por Virus ARN , Replicación Viral , Animales , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/virología , Inmunidad Innata/genética , Nodaviridae/fisiología , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Infecciones por Virus ARN/inmunología , Infecciones por Virus ARN/veterinaria , Lubina/inmunología , Filogenia , Regulación de la Expresión Génica/inmunología , Secuencia de Aminoácidos , Alineación de Secuencia/veterinaria , Enzimas Desubicuitinizantes/genética , Enzimas Desubicuitinizantes/inmunología , Perfilación de la Expresión Génica/veterinaria

RESUMEN

Inflammatory bowel disease (IBD), which include Crohn's disease (CD) and ulcerative colitis (UC), exhibits a complex multifactorial pathogenesis involving genetic susceptibility, imbalance of gut microbiota, mucosal immune disorder and environmental factors. Recent studies reported associations between ubiquitination and deubiquitination and the occurrence and development of inflammatory bowel disease. Ubiquitination modification, one of the most important types of post-translational modifications, is a multi-step enzymatic process involved in the regulation of various physiological processes of cells, including cell cycle progression, cell differentiation, apoptosis, and innate and adaptive immune responses. Alterations in ubiquitination and deubiquitination can lead to various diseases, including IBD. Here, we review the role of E3 ubiquitin ligases and deubiquitinases (DUBs) and their mediated ubiquitination and deubiquitination modifications in the pathogenesis of IBD. We highlight the importance of this type of posttranslational modification in the development of inflammation, and provide guidance for the future development of targeted therapeutics in IBD.

Asunto(s)

Enzimas Desubicuitinizantes/inmunología , Enfermedades Inflamatorias del Intestino/inmunología , Procesamiento Proteico-Postraduccional/inmunología , Transducción de Señal/inmunología , Ubiquitina-Proteína Ligasas/inmunología , Ubiquitinación/inmunología , Inmunidad Adaptativa/inmunología , Colitis Ulcerosa/enzimología , Colitis Ulcerosa/inmunología , Colitis Ulcerosa/metabolismo , Enfermedad de Crohn/enzimología , Enfermedad de Crohn/inmunología , Enfermedad de Crohn/metabolismo , Enzimas Desubicuitinizantes/metabolismo , Humanos , Inmunidad Innata/inmunología , Inflamación/enzimología , Inflamación/inmunología , Inflamación/metabolismo , Enfermedades Inflamatorias del Intestino/enzimología , Enfermedades Inflamatorias del Intestino/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo

RESUMEN

Viral infectious diseases pose a great challenge to human health around the world. Type I interferons (IFN-Is) function as the first line of host defense and thus play critical roles during virus infection by mediating the transcriptional induction of hundreds of genes. Nevertheless, overactive cytokine immune responses also cause autoimmune diseases, and thus, tight regulation of the innate immune response is needed to achieve viral clearance without causing excessive immune responses. Emerging studies have recently uncovered that the ubiquitin system, particularly deubiquitinating enzymes (DUBs), plays a critical role in regulating innate immune responses. In this review, we highlight recent advances on the diverse mechanisms of human DUBs implicated in IFN-I signaling. These DUBs function dynamically to calibrate host defenses against various virus infections by targeting hub proteins in the IFN-I signaling transduction pathway. We also present a future perspective on the roles of DUB-substrate interaction networks in innate antiviral activities, discuss the promises and challenges of DUB-based drug development, and identify the open questions that remain to be clarified. Our review provides a comprehensive description of DUBs, particularly their differential mechanisms that have evolved in the host to regulate IFN-I-signaling-mediated antiviral responses.

Asunto(s)

Enzimas Desubicuitinizantes/inmunología , Inmunidad Innata/inmunología , Interferón Tipo I/inmunología , Virosis/inmunología , Animales , Humanos , Transducción de Señal/inmunología

RESUMEN

Cancer immunotherapy has become an attractive approach of cancer treatment with tremendous success in treating various advanced malignancies. The development and clinical application of immune checkpoint inhibitors represent one of the most extraordinary accomplishments in cancer immunotherapy. In addition, considerable progress is being made in understanding the mechanism of antitumor immunity and characterizing novel targets for developing additional therapeutic approaches. One active area of investigation is protein ubiquitination, a post-translational mechanism of protein modification that regulates the function of diverse immune cells in antitumor immunity. Accumulating studies suggest that E3 ubiquitin ligases and deubiquitinases form a family of potential targets to be exploited for enhancing antitumor immunity in cancer immunotherapy.

Asunto(s)

Inhibidores de Puntos de Control Inmunológico/uso terapéutico , Inmunoterapia , Proteínas de Neoplasias/inmunología , Neoplasias , Ubiquitina/inmunología , Ubiquitinación/inmunología , Enzimas Desubicuitinizantes/inmunología , Humanos , Neoplasias/inmunología , Neoplasias/terapia , Ubiquitina-Proteína Ligasas/inmunología

RESUMEN

BACKGROUND: Very-early-onset inflammatory bowel disease (VEOIBD) is a chronic inflammatory disease of the gastrointestinal tract occurring during infancy or early childhood. NOD-like receptor family, pyrin domain containing 3 (NLRP3) inflammasome has emerged as a crucial regulator of intestinal homeostasis; however, whether NLRP3 variants may modify VEOIBD risk is unknown. OBJECTIVE: We sought to investigate whether and how a rare NLRP3 variant, found in 3 patients with gastrointestinal symptoms, contributes to VEOIBD development. METHODS: Whole-exome sequencing and bioinformatic analysis were performed to screen disease-associated NLRP3 variants from a cohort of children with VEOIBD. Inflammasome activation was determined in reconstituted HEK293T human embryonic kidney cells with NLRP3 inflammasome components, doxycycline-inducible NLRP3 macrophages, as well as PBMCs and biopsies from patients with NLRP3 variants. Pathogenesis of the variants was determined using a dextran sulfate sodium-induced acute colitis model. RESULTS: We identified a dominant gain-of-function missense variant of NLRP3, encoded by rs772009059 (R779C), in 3 patients with gastrointestinal symptoms. Functional analysis revealed that R779C increased NLRP3 inflammasome activation and pyroptosis in macrophages. This was mediated by enhanced deubiquitination of NLRP3 via binding with deubiquitinases BRCC3 and JOSD2, which are highly expressed in myeloid cells. In a dextran sulfate sodium-induced acute colitis model, NLRP3-R779C in hematopoietic cells resulted in more severe colitis, which can be ameliorated via knockdown of BRCC3 or JOSD2. CONCLUSIONS: BRCC3 and JOSD2 mediate NLRP3-R779C deubiquitination, which promotes NLRP3 inflammasome activation and the risk of developing VEOIBD.

Asunto(s)

Enfermedades Inflamatorias del Intestino , Mutación Missense , Proteína con Dominio Pirina 3 de la Familia NLR , Ubiquitinación , Edad de Inicio , Sustitución de Aminoácidos , Animales , Biopsia , Enzimas Desubicuitinizantes/inmunología , Femenino , Células HEK293 , Humanos , Lactante , Enfermedades Inflamatorias del Intestino/genética , Enfermedades Inflamatorias del Intestino/inmunología , Enfermedades Inflamatorias del Intestino/patología , Masculino , Ratones , Ratones Noqueados , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Factores de Riesgo , Células THP-1 , Secuenciación del Exoma

RESUMEN

BRCA1-BRCA2-containing complex subunit 3 (BRCC3), a Lys-63-specific deubiquitinase, is a member of the JAMM/MPN family of zinc metalloproteases. BRCC3 have been shown to promote the inflammasome activation by deubiquitinating NOD-like receptor containing pyrin domain 3 (NLRP3). We reported the involvement of neuronal inflammasome in Parkinson's Disease (PD), but the molecular mechanism remains unknown. In this study, we showed that BRCC3 expression was increased in PD models. Knock-down of BRCC3 with shRNA lentivirus decreased NLRP3 neuronal inflammasome. Interestingly, upregulating cyclin-dependent kinase 5 (Cdk5) increased the expression of BRCC3 in HEK293 cell, while inhibition of Cdk5 decreased the upregulated BRCC3 level in MPP+-induced PD cell model. The interaction between Cdk5 and BRCC3 was further confirmed by immunoprecipitation. Moreover, inhibition of Cdk5 suppressed the expression of NLRP3, pro-caspase-1, the adaptor molecule apoptosis-associated speck-like protein containing a CARD (ASC) and interleukin-1 beta (IL-1ß). Besides, inhibition of BRCC3 blocked the increased secretion of IL-1ß. Together, these results suggest that Cdk5-mediated BRCC3 expression may play a critical role in neuronal inflammation by regulating the NLRP3 inflammasome in PD.

Asunto(s)

Quinasa 5 Dependiente de la Ciclina/inmunología , Enzimas Desubicuitinizantes/inmunología , Inflamasomas/inmunología , Neuronas/inmunología , Enfermedad de Parkinson/inmunología , Animales , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/inmunología , Células Cultivadas , Enzimas Desubicuitinizantes/genética , Modelos Animales de Enfermedad , Células HEK293 , Humanos , Masculino , Ratones Endogámicos C57BL , Neuronas/patología , Enfermedad de Parkinson/genética , Enfermedad de Parkinson/patología , Ratas Sprague-Dawley , Regulación hacia Arriba

RESUMEN

In mammals, ∼100 deubiquitinases act on ∼20,000 intracellular ubiquitination sites. Deubiquitinases are commonly regarded as constitutively active, with limited regulatory and targeting capacity. The BRCA1-A and BRISC complexes serve in DNA double-strand break repair and immune signaling and contain the lysine-63 linkage-specific BRCC36 subunit that is functionalized by scaffold subunits ABRAXAS and ABRO1, respectively. The molecular basis underlying BRCA1-A and BRISC function is currently unknown. Here we show that in the BRCA1-A complex structure, ABRAXAS integrates the DNA repair protein RAP80 and provides a high-affinity binding site that sequesters the tumor suppressor BRCA1 away from the break site. In the BRISC structure, ABRO1 binds SHMT2α, a metabolic enzyme enabling cancer growth in hypoxic environments, which we find prevents BRCC36 from binding and cleaving ubiquitin chains. Our work explains modularity in the BRCC36 DUB family, with different adaptor subunits conferring diversified targeting and regulatory functions.

Asunto(s)

Proteína BRCA1/genética , Reparación del ADN/genética , Proteínas de Unión al ADN/genética , Enzimas Desubicuitinizantes/genética , Chaperonas de Histonas/genética , Neoplasias/genética , Sitios de Unión/genética , Proteínas Portadoras/genética , Núcleo Celular/genética , Núcleo Celular/inmunología , Citoplasma/genética , Citoplasma/inmunología , Roturas del ADN de Doble Cadena , Reparación del ADN/inmunología , Enzimas Desubicuitinizantes/inmunología , Células HeLa , Humanos , Inmunidad Celular/genética , Complejos Multiproteicos/química , Complejos Multiproteicos/genética , Neoplasias/inmunología , Proteínas Asociadas a Matriz Nuclear/genética , Unión Proteica/genética , Ubiquitina/genética , Proteasas Ubiquitina-Específicas/genética , Ubiquitinación/genética

Asunto(s)

Enfermedades Autoinmunes/inmunología , Enzimas Desubicuitinizantes/genética , Neoplasias/inmunología , Procesamiento Proteico-Postraduccional , Ubiquitina-Proteína Ligasas/genética , Animales , Enfermedades Autoinmunes/enzimología , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/patología , Enzimas Desubicuitinizantes/clasificación , Enzimas Desubicuitinizantes/inmunología , Humanos , Linfocitos/enzimología , Linfocitos/inmunología , Linfocitos/patología , Neoplasias/enzimología , Neoplasias/genética , Neoplasias/patología , Ubiquitina/genética , Ubiquitina/inmunología , Ubiquitina-Proteína Ligasas/clasificación , Ubiquitina-Proteína Ligasas/inmunología , Ubiquitinación

RESUMEN

T cells play critical roles in immune responses to pathogens, autoimmunity, and antitumor immunity. During the past few decades, increasing numbers of studies have demonstrated the significance of protein ubiquitination in T cell-mediated immunity. Several E3 ubiquitin ligases and deubiquitinases (DUBs) have been identified as either positive or negative regulators of T cell development and function. In this review, we mainly focus on the roles of DUBs (especially ubiquitin-specific proteases (USPs)) in modulating T cell differentiation and function, as well as the molecular mechanisms. Understanding how T cell development and function is regulated by ubiquitination and deubiquitination will provide novel strategies for treating infection, autoimmune diseases, and cancer.

Asunto(s)

Enfermedades Autoinmunes/inmunología , Enzimas Desubicuitinizantes/genética , Neoplasias/inmunología , Procesamiento Proteico-Postraduccional , Linfocitos T/inmunología , Ubiquitina-Proteína Ligasas/genética , Animales , Enfermedades Autoinmunes/enzimología , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/patología , Diferenciación Celular , Proliferación Celular , Enzimas Desubicuitinizantes/inmunología , Humanos , Tolerancia Inmunológica , Inmunidad Celular , Isoenzimas/genética , Isoenzimas/inmunología , Ratones , Neoplasias/enzimología , Neoplasias/genética , Neoplasias/patología , Transducción de Señal , Linfocitos T/enzimología , Linfocitos T/patología , Ubiquitina-Proteína Ligasas/inmunología , Ubiquitinación

RESUMEN

Host anti-viral innate-immune signalling pathways are regulated by a variety of post-translation modifications including ubiquitination, which is critical to regulate various signalling pathways for synthesis of anti-viral molecules. A homeostasis of host immune responses, induced due to viral infection and further ubiquitination, is maintained by the action of deubiquitinases (DUB). Infecting viruses utilize the process of deubiquitination for tricking host immune system wherein viral DUBs compete with host DUBs for inhibition of innate-immune anti-viral signalling pathways, which instead of maintaining an immune homeostasis bring about virus-mediated pathogenesis. This suggests that viruses co-evolve with their hosts to acquire similar machinery for tricking immune surveillance and establishing infection.

Asunto(s)

Enzimas Desubicuitinizantes/inmunología , Proteínas Virales/inmunología , Virosis/inmunología , Virus/enzimología , Animales , Enzimas Desubicuitinizantes/genética , Interacciones Huésped-Patógeno , Humanos , Evasión Inmune , Inmunidad Innata , Proteínas Virales/genética , Virosis/enzimología , Virosis/genética , Virosis/virología , Virus/genética , Virus/inmunología

RESUMEN

Cancer is a multifactorial disease and virus-mediated carcinogenesis is one of the crucial factors, which is poorly understood. Human cytomegalovirus (HCMV) is a herpesvirus and its components have been evidenced to be associated with cancer of different tissue origin. However, its role in cancer remains unknown. Here, we identified a conserved herpesviral tegument protein known as pUL48 of HCMV, encoding deubiquitinase enzyme, as having a key role in carcinogenesis. We show using deubiquitinase sufficient- and deficient-HCMV that HCMV deubiquitinase is a key in inducing enhanced cellular metabolic activity through upregulation of several anti-apoptotic genes and downregulation of several pro-apoptotic genes expression. Furthermore, HCMV deubiquitinase acquires pro-tumor functions by inhibiting PRR-mediated type I interferon via deubiquitination of TRAF6, TRAF3, IRAK1, IRF7 and STING. Taken together, our results suggest that HCMV infection may promote oncogenesis by inhibiting innate immunity of the host.

Asunto(s)

Carcinogénesis/genética , Enzimas Desubicuitinizantes/inmunología , Neoplasias/virología , Proteínas de la Matriz Viral/inmunología , Línea Celular Tumoral , Citomegalovirus/genética , Citomegalovirus/patogenicidad , Enzimas Desubicuitinizantes/genética , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Humanos , Inmunidad Innata/genética , Neoplasias/genética , Neoplasias/inmunología , Activación Transcripcional , Proteínas de la Matriz Viral/genética , Replicación Viral/genética

RESUMEN

Many infections and stress signals can rapidly activate the NLRP3 inflammasome to elicit robust inflammatory responses. This activation requires a priming step, which is thought to be mainly for upregulating NLRP3 transcription. However, recent studies report that the NLRP3 inflammasome can be activated independently of transcription, suggesting that the priming process has unknown essential regulatory steps. Here, we report that JNK1-mediated NLRP3 phosphorylation at S194 is a critical priming event and is essential for NLRP3 inflammasome activation. We show that NLRP3 inflammasome activation is disrupted in NLRP3-S194A knockin mice. JNK1-mediated NLRP3 S194 phosphorylation is critical for NLRP3 deubiquitination and facilitates its self-association and the subsequent inflammasome assembly. Importantly, we demonstrate that blocking S194 phosphorylation prevents NLRP3 inflammasome activation in cryopyrin-associated periodic syndromes (CAPS). Thus, our study reveals a key priming molecular event that is a prerequisite for NLRP3 inflammasome activation. Inhibiting NLRP3 phosphorylation could be an effective treatment for NLRP3-related diseases.

Asunto(s)

Inflamasomas/genética , Macrófagos/inmunología , Proteína Quinasa 8 Activada por Mitógenos/genética , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Choque Séptico/genética , Secuencia de Aminoácidos , Animales , Enzimas Desubicuitinizantes/genética , Enzimas Desubicuitinizantes/inmunología , Escherichia coli/química , Femenino , Regulación de la Expresión Génica , Células HEK293 , Humanos , Inflamasomas/inmunología , Lipopolisacáridos/farmacología , Macrófagos/patología , Masculino , Ratones , Ratones Transgénicos , Proteína Quinasa 8 Activada por Mitógenos/inmunología , Proteína con Dominio Pirina 3 de la Familia NLR/deficiencia , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Fosforilación , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Choque Séptico/inducido químicamente , Choque Séptico/mortalidad , Choque Séptico/patología , Transducción de Señal , Análisis de Supervivencia

RESUMEN

Understanding of molecular mechanisms that drive Waldenstrom macroglobulinemia (WM) cell survival are rapidly evolving. This review briefly highlights emerging "WM-relevant" targets; for which therapeutic strategies are currently being investigated in preclinical and clinical studies. With the discovery of MYD88L265P signaling and remarkable activity of ibrutinib in WM, other targets within the B-cell receptor pathway are now being focused on for therapeutic intervention. Additional targets which play a role in WM cell survival include TLR7, 8 and 9, proteasome-associated deubiquitinating enzymes (USP14 and UCHL5), XPO1/CRM1 and AURKA. New drugs for established targets are also discussed. Lastly, we spotlight 3 highly innovative WM-specific therapies: MYD88 peptide inhibitors, MYD88L265P-directed immune activation and CD19-directed chimeric antigen receptor T-cell therapy, which are in various stages of development. Indeed, treatment of WM is poised to undergo a paradigm shift in the coming years towards highly disease-driven and more personalized therapeutic modalities with curative intent.

Asunto(s)

Antineoplásicos/uso terapéutico , Tratamiento Basado en Trasplante de Células y Tejidos , Transfusión de Linfocitos , Receptores de Antígenos de Linfocitos T , Proteínas Recombinantes de Fusión , Macroglobulinemia de Waldenström/terapia , Sustitución de Aminoácidos , Animales , Antígenos CD19/genética , Antígenos CD19/inmunología , Aurora Quinasa A/antagonistas & inhibidores , Aurora Quinasa A/genética , Aurora Quinasa A/inmunología , Enzimas Desubicuitinizantes/antagonistas & inhibidores , Enzimas Desubicuitinizantes/genética , Enzimas Desubicuitinizantes/inmunología , Humanos , Mutación Missense , Factor 88 de Diferenciación Mieloide/antagonistas & inhibidores , Factor 88 de Diferenciación Mieloide/genética , Factor 88 de Diferenciación Mieloide/inmunología , Receptores Toll-Like/antagonistas & inhibidores , Receptores Toll-Like/genética , Receptores Toll-Like/inmunología , Macroglobulinemia de Waldenström/genética , Macroglobulinemia de Waldenström/inmunología