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1.
Immunity ; 53(6): 1151-1167.e6, 2020 12 15.
Artículo en Inglés | MEDLINE | ID: mdl-33159853

RESUMEN

Establishment of B-lineage-specific gene expression requires the binding of transcription factors to inaccessible chromatin of progenitors. The transcription factor EBF1 can bind genomic regions prior to the detection of chromatin accessibility in a manner dependent on EBF1's C-terminal domain (CTD) and independent of cooperating transcription factors. Here, we studied the mechanism whereby the CTD enables this pioneering function. The CTD of EBF1 was dispensable for initial chromatin targeting but stabilized occupancy via recruitment of the chromatin remodeler Brg1. We found that the CTD harbors a prion-like domain (PLD) with an ability of liquid-liquid phase separation, which was enhanced by interaction of EBF1 with the RNA-binding protein FUS. Brg1 also partitioned into phase-separated FUS condensates and coincided with EBF1 and FUS foci in pro-B cells. Heterologous PLDs conferred pioneering function on EBF1ΔCTD. Thus, the phase separation ability of EBF1 facilitates Brg1-mediated chromatin opening and the transition of naive progenitor chromatin to B-lineage-committed chromatin.


Asunto(s)
Linfocitos B/metabolismo , Cromatina/metabolismo , Priones/química , Transactivadores/metabolismo , Secuencia de Aminoácidos , Linfocitos B/citología , ADN Helicasas/metabolismo , Humanos , Proteínas Nucleares/metabolismo , Transición de Fase , Células Precursoras de Linfocitos B/citología , Células Precursoras de Linfocitos B/metabolismo , Dominios Proteicos , Proteína FUS de Unión a ARN/metabolismo , Proteínas de Unión al ARN/metabolismo , Transactivadores/química , Factores de Transcripción/metabolismo
2.
Genes Dev ; 30(20): 2310-2324, 2016 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-27807034

RESUMEN

Transcription factor EBF1 (early B-cell factor 1) regulates early B-cell differentiation by poising or activating lineage-specific genes and repressing genes associated with alternative cell fates. To identify proteins that regulate the diverse functions of EBF1, we used SILAC (stable isotope labeling by amino acids in cell culture)-based mass spectrometry of proteins associated with endogenous EBF1 in pro-B cells. This analysis identified most components of the multifunctional CCR4-NOT complex, which regulates transcription and mRNA degradation. CNOT3 interacts with EBF1, and we identified histidine 240 in EBF1 as a critical residue for this interaction. Complementation of Ebf1-/- progenitors with EBF1H240A revealed a partial block of pro-B-cell differentiation and altered expression of specific EBF1 target genes that show either reduced transcription or increased mRNA stability. Most deregulated EBF1 target genes show normal occupancy by EBF1H240A, but we also detected genes with altered occupancy, suggesting that the CCR4-NOT complex affects multiple activities of EBF1. Mice with conditional Cnot3 inactivation recapitulate the block of early B-cell differentiation, which we found to be associated with an impaired autoregulation of Ebf1 and reduced expression of pre-B-cell receptor components. Thus, the interaction of the CCR4-NOT complex with EBF1 diversifies the function of EBF1 in a context-dependent manner and may coordinate transcriptional and post-transcriptional gene regulation.


Asunto(s)
Linfocitos B/fisiología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Regulación de la Expresión Génica/genética , Proteínas de Homeodominio/metabolismo , Linfopoyesis/genética , Proteínas Nucleares/metabolismo , Estabilidad del ARN/genética , Factores de Transcripción/metabolismo , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Diferenciación Celular/genética , Silenciador del Gen , Células HEK293 , Proteínas de Homeodominio/genética , Humanos , Ratones , Chaperonas Moleculares/genética , Mutación , Proteínas Nucleares/genética , Unión Proteica , Factores de Transcripción/genética , Transgenes
3.
Proc Natl Acad Sci U S A ; 110(30): 12331-6, 2013 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-23836662

RESUMEN

The circadian clock gene Period2 (PER2) has been suggested to be a tumor suppressor. However, detailed mechanistic evidence has not been provided to support this hypothesis. We found that loss of PER2 enhanced invasion and activated expression of epithelial-mesenchymal transition (EMT) genes including TWIST1, SLUG, and SNAIL. This finding was corroborated by clinical observation that PER2 down-regulation was associated with poor prognosis in breast cancer patients. We further demonstrated that PER2 served as a transcriptional corepressor, which recruited polycomb proteins EZH2 and SUZ12 as well as HDAC2 to octamer transcription factor 1 (OCT1) (POU2F1) binding sites of the TWIST1 and SLUG promoters to repress expression of these EMT genes. Hypoxia, a condition commonly observed in tumors, caused PER2 degradation and disrupted the PER2 repressor complex, leading to activation of EMT gene expression. This result was further supported by clinical data showing a significant negative correlation between hypoxia and PER2. Thus, our findings clearly demonstrate the tumor suppression function of PER2 and elucidate a pathway by which hypoxia promotes EMT via degradation of PER2.


Asunto(s)
Neoplasias de la Mama/metabolismo , Transición Epitelial-Mesenquimal , Regulación de la Expresión Génica/genética , Hipoxia/genética , Transportador 1 de Catión Orgánico/fisiología , Proteínas Circadianas Period/genética , Neoplasias de la Mama/patología , Línea Celular Tumoral , Regulación hacia Abajo , Femenino , Humanos , Regiones Promotoras Genéticas , Procesamiento Proteico-Postraduccional , Regulación hacia Arriba/genética
4.
Free Radic Biol Med ; 165: 368-384, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33460768

RESUMEN

Emerging evidences implicate the contribution of ROS to T cell activation and signaling. The tyrosine kinase, ζ-chain-associated protein of 70 kDa (ZAP70), is essential for T cell development and activation. However, it remains elusive whether a direct redox regulation affects ZAP70 activity upon TCR stimulation. Here, we show that deficiency of non-selenocysteine containing phospholipid hydroperoxide glutathione peroxidase (NPGPx), a redox sensor, results in T cell hyperproliferation and elevated cytokine productions. T cell-specific NPGPx-knockout mice reveal enhanced T-dependent humoral responses and are susceptible to experimental autoimmune encephalomyelitis (EAE). Through proteomic approaches, ZAP70 is identified as the key interacting protein of NPGPx through disulfide bonding. NPGPx is activated by ROS generated from TCR stimulation, and modulates ZAP70 activity through redox switching to reduce ZAP70 recruitment to TCR/CD3 complex in membrane lipid raft, therefore subduing TCR responses. These results reveal a delicate redox mechanism that NPGPx serves as a modulator to curb ZAP70 functions in maintaining T cell homeostasis.


Asunto(s)
Proteómica , Linfocitos T , Animales , Homeostasis , Ratones , Oxidación-Reducción , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/metabolismo , Transducción de Señal , Linfocitos T/metabolismo
5.
Oncol Rep ; 40(3): 1795-1802, 2018 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-30015941

RESUMEN

Angiogenesis is crucial for the progression of esophageal squamous cell carcinoma (ESCC). Anti­angiogenesis by targeting important molecules has been considered as one of the most promising and efficient strategy for cancer therapy. Recent studies have demonstrated that the IQ­domain GTPase activating protein 1 (IQGAP1) plays critical roles in tumorigenesis and cancer progression. We previously reported that IQGAP1 is overexpressed in ESCC, and IQGAP1 knockdown can decrease cell proliferation and metastasis ability in vitro and in vivo. However, the effects of IQGAP1 on the angiogenesis of ESCC and its underlying mechanisms remain unknown. In the present study, we found that IQGAP1 overexpression promoted tumor angiogenesis confirmed by human umbilical vascular endothelial cell (HUVEC) tube formation in vitro and chicken embryo chorioallantoic membrane (CAM) assay in vivo. Moreover, IQGAP1 overexpression in ESCC cells increased expression of vascular endothelial growth factor (VEGF) and phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2). Meanwhile, we found that levels of AKT and ERK phosphorylation were upregulated in IQGAP1­overexpressing cells. Importantly, IQGAP1­knockdown cells showed the opposing results. Furthermore, AKT and ERK inhibitors not only significantly decreased VEGF expression and VEGFR2 phosphorylation in IQGAP1­overexpressing cells, but also abolished the pro­angiogenic effect of IQGAP1 overexpression on angiogenesis in the HUVEC tube formation and chicken embryo CAM assay. Taken together, this evidence confirms that IQGAP1 overexpression promotes tumor angiogenesis via the AKT and ERK­mediated VEGF­VEGFR2 signaling pathway in ESCC, and IQGAP1 may be an attractive therapeutic target for cancer anti­angiogenesis treatment.


Asunto(s)
Neoplasias Esofágicas/irrigación sanguínea , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Neovascularización Patológica , Proteínas Proto-Oncogénicas c-akt/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , Proteínas Activadoras de ras GTPasa/metabolismo , Apoptosis , Biomarcadores de Tumor/metabolismo , Carcinoma de Células Escamosas/irrigación sanguínea , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Proliferación Celular , Membrana Corioalantoides/metabolismo , Membrana Corioalantoides/patología , Neoplasias Esofágicas/metabolismo , Neoplasias Esofágicas/patología , Humanos , Fosforilación , Células Tumorales Cultivadas
7.
Nat Genet ; 46(2): 176-181, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24362818

RESUMEN

Follicular lymphoma is an incurable malignancy, with transformation to an aggressive subtype representing a critical event during disease progression. Here we performed whole-genome or whole-exome sequencing on 10 follicular lymphoma-transformed follicular lymphoma pairs followed by deep sequencing of 28 genes in an extension cohort, and we report the key events and evolutionary processes governing tumor initiation and transformation. Tumor evolution occurred through either a 'rich' or 'sparse' ancestral common progenitor clone (CPC). We identified recurrent mutations in linker histone, JAK-STAT signaling, NF-κB signaling and B cell developmental genes. Longitudinal analyses identified early driver mutations in chromatin regulator genes (CREBBP, EZH2 and KMT2D (MLL2)), whereas mutations in EBF1 and regulators of NF-κB signaling (MYD88 and TNFAIP3) were gained at transformation. Collectively, this study provides new insights into the genetic basis of follicular lymphoma and the clonal dynamics of transformation and suggests that personalizing therapies to target key genetic alterations in the CPC represents an attractive therapeutic strategy.


Asunto(s)
Transformación Celular Neoplásica/genética , Progresión de la Enfermedad , Genómica/métodos , Linfoma Folicular/genética , Linfoma Folicular/fisiopatología , Secuencia de Bases , Proteína de Unión a CREB/genética , Análisis por Conglomerados , Estudios de Cohortes , Proteínas de Unión al ADN/genética , Proteína Potenciadora del Homólogo Zeste 2 , Exoma/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Histonas/genética , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Mutagénesis , Mutación/genética , Factor 88 de Diferenciación Mieloide/genética , Proteínas de Neoplasias/genética , Proteínas Nucleares/genética , Filogenia , Complejo Represivo Polycomb 2/genética , Polimorfismo de Nucleótido Simple/genética , Análisis de Secuencia de ADN , Transactivadores/genética , Proteína 3 Inducida por el Factor de Necrosis Tumoral alfa
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