Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 5 de 5
Filtrar
Más filtros

Banco de datos
Tipo del documento
País de afiliación
Intervalo de año de publicación
1.
Nat Immunol ; 17(3): 250-8, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26642356

RESUMEN

The NLRP3 inflammasome responds to microbes and danger signals by processing and activating proinflammatory cytokines, including interleukin 1ß (IL-1ß) and IL-18. We found here that activation of the NLRP3 inflammasome was restricted to interphase of the cell cycle by NEK7, a serine-threonine kinase previously linked to mitosis. Activation of the NLRP3 inflammasome required NEK7, which bound to the leucine-rich repeat domain of NLRP3 in a kinase-independent manner downstream of the induction of mitochondrial reactive oxygen species (ROS). This interaction was necessary for the formation of a complex containing NLRP3 and the adaptor ASC, oligomerization of ASC and activation of caspase-1. NEK7 promoted the NLRP3-dependent cellular inflammatory response to intraperitoneal challenge with monosodium urate and the development of experimental autoimmune encephalitis in mice. Our findings suggest that NEK7 serves as a cellular switch that enforces mutual exclusivity of the inflammasome response and cell division.


Asunto(s)
Proteínas Portadoras/inmunología , Macrófagos/inmunología , Mitosis/inmunología , Proteínas Serina-Treonina Quinasas/inmunología , Animales , Apoptosis , Proteínas Reguladoras de la Apoptosis , Proteínas Adaptadoras de Señalización CARD , Proteínas Portadoras/genética , Caspasa 1 , Cromatografía en Gel , Ensayo de Unidades Formadoras de Colonias , Citocinas , Proteínas del Citoesqueleto , Células Dendríticas , Encefalomielitis Autoinmune Experimental/inmunología , Femenino , Citometría de Flujo , Células HEK293 , Humanos , Inmunoprecipitación , Técnicas In Vitro , Inflamasomas/genética , Inflamasomas/inmunología , Macrófagos Peritoneales/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mitocondrias/metabolismo , Monocitos , Quinasas Relacionadas con NIMA , Proteína con Dominio Pirina 3 de la Familia NLR , Proteínas Serina-Treonina Quinasas/genética , Especies Reactivas de Oxígeno , Médula Espinal/inmunología
2.
Proc Natl Acad Sci U S A ; 112(5): E440-9, 2015 Feb 03.
Artículo en Inglés | MEDLINE | ID: mdl-25605905

RESUMEN

With the wide availability of massively parallel sequencing technologies, genetic mapping has become the rate limiting step in mammalian forward genetics. Here we introduce a method for real-time identification of N-ethyl-N-nitrosourea-induced mutations that cause phenotypes in mice. All mutations are identified by whole exome G1 progenitor sequencing and their zygosity is established in G2/G3 mice before phenotypic assessment. Quantitative and qualitative traits, including lethal effects, in single or multiple combined pedigrees are then analyzed with Linkage Analyzer, a software program that detects significant linkage between individual mutations and aberrant phenotypic scores and presents processed data as Manhattan plots. As multiple alleles of genes are acquired through mutagenesis, pooled "superpedigrees" are created to analyze the effects. Our method is distinguished from conventional forward genetic methods because it permits (1) unbiased declaration of mappable phenotypes, including those that are incompletely penetrant (2), automated identification of causative mutations concurrent with phenotypic screening, without the need to outcross mutant mice to another strain and backcross them, and (3) exclusion of genes not involved in phenotypes of interest. We validated our approach and Linkage Analyzer for the identification of 47 mutations in 45 previously known genes causative for adaptive immune phenotypes; our analysis also implicated 474 genes not previously associated with immune function. The method described here permits forward genetic analysis in mice, limited only by the rates of mutant production and screening.


Asunto(s)
Mutación Puntual , Alelos , Animales , Femenino , Genes Letales , Ligamiento Genético , Masculino , Ratones , Linaje , Fenotipo , Sitios de Carácter Cuantitativo
3.
Nat Neurosci ; 18(8): 1094-100, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-26192746

RESUMEN

The cAMP and cAMP-dependent protein kinase A (PKA) signaling cascade is a ubiquitous pathway acting downstream of multiple neuromodulators. We found that the phosphorylation of phosphodiesterase-4 (PDE4) by cyclin-dependent protein kinase 5 (Cdk5) facilitated cAMP degradation and homeostasis of cAMP/PKA signaling. In mice, loss of Cdk5 throughout the forebrain elevated cAMP levels and increased PKA activity in striatal neurons, and altered behavioral responses to acute or chronic stressors. Ventral striatum- or D1 dopamine receptor-specific conditional knockout of Cdk5, or ventral striatum infusion of a small interfering peptide that selectively targeted the regulation of PDE4 by Cdk5, produced analogous effects on stress-induced behavioral responses. Together, our results demonstrate that altering cAMP signaling in medium spiny neurons of the ventral striatum can effectively modulate stress-induced behavioral states. We propose that targeting the Cdk5 regulation of PDE4 could be a new therapeutic approach for clinical conditions associated with stress, such as depression.


Asunto(s)
Conducta Animal/fisiología , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 4/metabolismo , Quinasa 5 Dependiente de la Ciclina/metabolismo , Transducción de Señal/fisiología , Estrés Psicológico/metabolismo , Estriado Ventral/metabolismo , Animales , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados
4.
Science ; 346(6216): 1486-92, 2014 Dec 19.
Artículo en Inglés | MEDLINE | ID: mdl-25525240

RESUMEN

Multivalent molecules with repetitive structures including bacterial capsular polysaccharides and viral capsids elicit antibody responses through B cell receptor (BCR) crosslinking in the absence of T cell help. We report that immunization with these T cell-independent type 2 (TI-2) antigens causes up-regulation of endogenous retrovirus (ERV) RNAs in antigen-specific mouse B cells. These RNAs are detected via a mitochondrial antiviral signaling protein (MAVS)-dependent RNA sensing pathway or reverse-transcribed and detected via the cGAS-cGAMP-STING pathway, triggering a second, sustained wave of signaling that promotes specific immunoglobulin M production. Deficiency of both MAVS and cGAS, or treatment of MAVS-deficient mice with reverse transcriptase inhibitors, dramatically inhibits TI-2 antibody responses. These findings suggest that ERV and two innate sensing pathways that detect them are integral components of the TI-2 B cell signaling apparatus.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/inmunología , Antígenos T-Independientes/inmunología , Linfocitos B/inmunología , Retrovirus Endógenos/inmunología , Nucleotidiltransferasas/inmunología , ARN Viral/inmunología , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Formación de Anticuerpos , Citosol/inmunología , ADN/inmunología , Retrovirus Endógenos/genética , Activación de Linfocitos , Proteínas de la Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Nucleótidos Cíclicos/inmunología , Nucleotidiltransferasas/genética , ARN Viral/genética , Transcripción Genética
5.
Science ; 321(5892): 1078-80, 2008 Aug 22.
Artículo en Inglés | MEDLINE | ID: mdl-18719281

RESUMEN

Many bacterial pathogens rely on a conserved membrane histidine sensor kinase, QseC, to respond to host adrenergic signaling molecules and bacterial signals in order to promote the expression of virulence factors. Using a high-throughput screen, we identified a small molecule, LED209, that inhibits the binding of signals to QseC, preventing its autophosphorylation and consequently inhibiting QseC-mediated activation of virulence gene expression. LED209 is not toxic and does not inhibit pathogen growth; however, this compound markedly inhibits the virulence of several pathogens in vitro and in vivo in animals. Inhibition of signaling offers a strategy for the development of broad-spectrum antimicrobial drugs.


Asunto(s)
Antibacterianos/farmacología , Escherichia coli Enterohemorrágica/patogenicidad , Proteínas de Escherichia coli/metabolismo , Francisella tularensis/patogenicidad , Infecciones por Bacterias Gramnegativas/tratamiento farmacológico , Proteínas Quinasas/metabolismo , Salmonella typhimurium/patogenicidad , Sulfonamidas/farmacología , Animales , Antibacterianos/administración & dosificación , Antibacterianos/uso terapéutico , Escherichia coli Enterohemorrágica/efectos de los fármacos , Escherichia coli Enterohemorrágica/genética , Escherichia coli Enterohemorrágica/metabolismo , Infecciones por Escherichia coli/tratamiento farmacológico , Proteínas de Escherichia coli/antagonistas & inhibidores , Proteínas de Escherichia coli/genética , Francisella tularensis/efectos de los fármacos , Francisella tularensis/genética , Francisella tularensis/metabolismo , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Histidina Quinasa , Ratones , Norepinefrina/metabolismo , Fosforilación , Proteínas Quinasas/genética , Conejos , Salmonelosis Animal/tratamiento farmacológico , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genética , Salmonella typhimurium/metabolismo , Transducción de Señal/efectos de los fármacos , Bibliotecas de Moléculas Pequeñas , Sulfonamidas/administración & dosificación , Sulfonamidas/química , Sulfonamidas/uso terapéutico , Tularemia/tratamiento farmacológico , Factores de Virulencia/genética
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA