RESUMEN
Transcription is extremely important for cellular processes but can be hindered by RNA polymerase II (RNAPII) pausing and stalling. Cockayne syndrome protein B (CSB) promotes the progression of paused RNAPII or initiates transcription-coupled nucleotide excision repair (TC-NER) to remove stalled RNAPII. However, the specific mechanism by which CSB initiates TC-NER upon damage remains unclear. In this study, we identified the indispensable role of the ARK2N-CK2 complex in the CSB-mediated initiation of TC-NER. The ARK2N-CK2 complex is recruited to damage sites through CSB and then phosphorylates CSB. Phosphorylation of CSB enhances its binding to stalled RNAPII, prolonging the association of CSB with chromatin and promoting CSA-mediated ubiquitination of stalled RNAPII. Consistent with this finding, Ark2n-/- mice exhibit a phenotype resembling Cockayne syndrome. These findings shed light on the pivotal role of the ARK2N-CK2 complex in governing the fate of RNAPII through CSB, bridging a critical gap necessary for initiating TC-NER.
Asunto(s)
Síndrome de Cockayne , ADN Helicasas , Enzimas Reparadoras del ADN , Reparación del ADN , Proteínas de Unión a Poli-ADP-Ribosa , ARN Polimerasa II , Enzimas Reparadoras del ADN/metabolismo , Enzimas Reparadoras del ADN/genética , ARN Polimerasa II/metabolismo , ARN Polimerasa II/genética , Proteínas de Unión a Poli-ADP-Ribosa/metabolismo , Proteínas de Unión a Poli-ADP-Ribosa/genética , Humanos , Animales , Ratones , ADN Helicasas/metabolismo , ADN Helicasas/genética , Síndrome de Cockayne/genética , Síndrome de Cockayne/metabolismo , Transcripción Genética , Fosforilación , Quinasa de la Caseína II/metabolismo , Quinasa de la Caseína II/genética , Ratones Noqueados , Daño del ADN , ATPasas Asociadas con Actividades Celulares Diversas/metabolismo , ATPasas Asociadas con Actividades Celulares Diversas/genética , Cromatina/metabolismo , Ubiquitinación , Reparación por EscisiónRESUMEN
Tandem DNA repeats are often organized into heterochromatin that is crucial for genome organization and stability. Recent studies revealed that individual repeats within tandem DNA repeats can behave very differently. How DNA repeats are assembled into distinct heterochromatin structures remains poorly understood. Here, we developed a genome-wide genetic screen using a reporter gene at different units in a repeat array. This screen led to identification of a conserved protein Rex1BD required for heterochromatin silencing. Our structural analysis revealed that Rex1BD forms a four-helix bundle structure with a distinct charged electrostatic surface. Mechanistically, Rex1BD facilitates the recruitment of Clr6 histone deacetylase (HDAC) by interacting with histones. Interestingly, Rex1BD also interacts with the 14-3-3 protein Rad25, which is responsible for recruiting the RITS (RNA-induced transcriptional silencing) complex to DNA repeats. Our results suggest that coordinated action of Rex1BD and Rad25 mediates formation of distinct heterochromatin structure at DNA repeats via linking RNAi and HDAC pathways.
Asunto(s)
Proteínas de Schizosaccharomyces pombe , Schizosaccharomyces , Interferencia de ARN , Heterocromatina/genética , Heterocromatina/metabolismo , Proteínas de Schizosaccharomyces pombe/metabolismo , Schizosaccharomyces/metabolismo , Proteínas 14-3-3/genética , Proteínas 14-3-3/metabolismo , Histona Desacetilasas/metabolismo , ADN/metabolismo , Secuencias Repetidas en TándemRESUMEN
BACKGROUND AND AIMS: Bile acids trigger a hepatic inflammatory response, causing cholestatic liver injury. Runt-related transcription factor-1 (RUNX1), primarily known as a master modulator in hematopoiesis, plays a pivotal role in mediating inflammatory responses. However, RUNX1 in hepatocytes is poorly characterized, and its role in cholestasis is unclear. Herein, we aimed to investigate the role of hepatic RUNX1 and its underlying mechanisms in cholestasis. APPROACH AND RESULTS: Hepatic expression of RUNX1 was examined in cholestatic patients and mouse models. Mice with liver-specific ablation of Runx1 were generated. Bile duct ligation and 1% cholic acid diet were used to induce cholestasis in mice. Primary mouse hepatocytes and the human hepatoma PLC/RPF/5- ASBT cell line were used for mechanistic studies. Hepatic RUNX1 mRNA and protein levels were markedly increased in cholestatic patients and mice. Liver-specific deletion of Runx1 aggravated inflammation and liver injury in cholestatic mice induced by bile duct ligation or 1% cholic acid feeding. Mechanistic studies indicated that elevated bile acids stimulated RUNX1 expression by activating the RUNX1 -P2 promoter through JAK/STAT3 signaling. Increased RUNX1 is directly bound to the promotor region of inflammatory chemokines, including CCL2 and CXCL2 , and transcriptionally repressed their expression in hepatocytes, leading to attenuation of liver inflammatory response. Blocking the JAK signaling or STAT3 phosphorylation completely abolished RUNX1 repression of bile acid-induced CCL2 and CXCL2 in hepatocytes. CONCLUSIONS: This study has gained initial evidence establishing the functional role of hepatocyte RUNX1 in alleviating liver inflammation during cholestasis through JAK/STAT3 signaling. Modulating hepatic RUNX1 activity could be a new therapeutic target for cholestasis.
Asunto(s)
Ácidos y Sales Biliares , Colestasis , Inflamación , Animales , Humanos , Ratones , Ácidos y Sales Biliares/efectos adversos , Ácidos y Sales Biliares/metabolismo , Colestasis/etiología , Colestasis/metabolismo , Ácidos Cólicos/efectos adversos , Ácidos Cólicos/farmacología , Subunidad alfa 2 del Factor de Unión al Sitio Principal/metabolismo , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Inflamación/etiología , Inflamación/genética , Inflamación/metabolismo , Hígado/metabolismo , Factor de Transcripción STAT3/metabolismoRESUMEN
Surface plasmons have robust and strong confinement to the light field which is beneficial for the light-matter interaction. Surface plasmon amplification by stimulated emission of radiation (SPACER) has the potential to be integrated on the semiconductor chip as a compact coherent light source, which can play an important role in further extension of Moore's law. In this study, we demonstrate the localized surface plasmon lasing at room temperature in the communication band using metallic nanoholes as the plasmonic nanocavity and InP nanowires as the gain medium. Optimizing laser performance has been demonstrated by coupling between two metallic nanoholes which adds another degree of freedom for manipulating the lasing properties. Our plasmonic nanolasers exhibit lower power consumption, smaller mode volumes, and higher spontaneous emission coupling factors due to enhanced light-matter interactions, which are very promising in the applications of high-density sensing and photonic integrated circuits.
RESUMEN
Photochromism is the heart of photochromic fluorescent proteins. Excited-state proton transfer (ESPT) is the major cause of photochromism for the green fluorescent protein (GFP) and Z-E photoisomerization through τ-torsion is the major cause of photochromism for Dronpa (a GFP mutant). In this article, s-E-1 opens a third type of photochromism for GFP chromophore derivatives, which involves light-driven φ-torsion with no τ-torsion, followed by excited-state intramolecular proton transfer (ESIPT), and is gated by environmental polarity. Since s-E-1 does not follow Z-E photoisomerization through τ-torsion but undergoes light-driven φ-torsion, which involves equilibration of the excited-state rotamers, it is clearly against the NEER (Non-Equilibration of Excited-state Rotamers) principle.
RESUMEN
Synthesis of C6+ higher alcohols from readily-accessible aqueous ethanol is an alternative route of great potential for blending-fuel, plasticizer, surfactant and medicine precursors, but the direct coupling of aqueous ethanol to C6+ higher alcohols is still challenging. Herein, the alkali carbonate induced N-doping of a NiSn@NC catalyst was achieved by a facile gel-carbonization strategy, and the effect of alkali salt inductors was examined for the direct coupling of 50 wt% aqueous ethanol. Noteworthily, C6+ higher alcohol selectivity of 61.9% with 57.1% ethanol conversion was achieved for the first time over the NiSn@NC-Na2CO3-1/9 catalyst, which broke the step-growth carbon distribution of ethanol coupling to higher alcohols. The inductive effect of alkali carbonate for the N doped graphite structure from the NO3- precursor was revealed. Electron transfer from Ni to the pyridine N doped graphite layer is enhanced, thus elevating the Ni-4s band center, which lowers the dehydrogenation barrier of the alcohol substrate and further improves the C6+OH selectivity. The catalyst reusability was also examined. This work gained new insight into the selective synthesis of high-carbon value-added chemicals from C-C coupling of aqueous ethanol.
Asunto(s)
Etanol , Grafito , Álcalis , Carbonatos , CarbonoRESUMEN
Zearalenone (ZEN), one of the most common mycotoxins in cereals, poses a severe health risk to humans. In this study, electrochemical oxidation and reduction degraded ZEN in solution completely within 8 min and 20 min. The structure of ZEN products was elucidated by mass spectrometry (MS), and their toxicity was evaluated by ECOSAR software and cytotoxicity assay. From simulation, electrochemical oxidation products had lower acute and chronic toxicity, and the product at 9.0 V is not harmful (LC50/EC50 greater than 100 mg/L, ChV greater than 10 mg/L). CCK-8 assay further confirmed their less cytotoxicity. To our surprise, LC50, EC50, and ChVs of all electrochemical reduction products were lower than 1 mg/L, and cell viabilities were less than ZEN, meaning the higher toxicity of electrochemical reduction products. On this Basis, electrochemical oxidation was applied in ZEN contaminated wheat with a degradation rate of 92.32 ± 2.37%, indicating its potential to degrade ZEN practically.
Asunto(s)
Micotoxinas , Zearalenona , Humanos , Zearalenona/análisis , Micotoxinas/análisis , Grano Comestible/química , Espectrometría de Masas , Dosificación Letal MedianaRESUMEN
Tumor necrosis factor receptor superfamily member-12A (TNFRSF12A) plays a critical role in inflammation and cell death. It is expressed in multiple tissues yet extremely low in normal liver. To date, little is known about its role in cholestasis. Therefore, we sought to delineate the role of TNFRSF12A in cholestasis and its underlying mechanisms. Human liver tissues were collected from patients with obstructive cholestasis (OC) or primary biliary cholangitis (PBC). Tnfrsf12a knockout (KO) mice were generated. Cholestasis was induced by bile-duct ligation (BDL) or 0.1% 5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-feeding. Human hepatoma PLC/PRF/5-ASBT and THP1 cell lines or primary mouse hepatocytes were used for mechanistic studies. Hepatic TNFRSF12A expression was markedly increased in OC or PBC patients. Genetic ablation of Tnfrsf12a in BDL- and 0.1%DDC-induced cholestatic mice significantly attenuated cholestatic liver injury with remarkable reduction of hepatocyte pyroptosis but without changing scores of necroptosis and apoptosis. Morphological features of hepatocyte pyroptosis and increased levels of pyroptosis-related proteins, NLRP3, cleaved-Caspase-1, and cleaved-GSDMD in OC patients and BDL-mice confirmed this observation. Further mechanistic studies revealed that bile acids (BAs) induced TNFRSF12A expression by enhancing the transcription factor c-JUN binding to the TNFRSF12A promoter and subsequently initiated hepatocyte pyroptosis by the NFκB/Caspase-1/GSDMD signaling. Interestingly, TWEAK, a typical ligand of TNFRSF12A, secreted by infiltrated macrophages in cholestatic livers, enhanced TNFRSF12A-induced hepatocyte pyroptosis. Taken together, we report, for the first time, that hepatic TNFRSF12A is dramatically increased in human cholestasis. Deletion of TNFRSF12A inhibits BAs-induced hepatocyte pyroptosis through the NFκB/Caspase-1/GSDMD signaling and thereby ameliorates cholestatic liver injury. As such, targeting TNFRSF12A could be a promising approach to treating cholestasis.
RESUMEN
Coating and cross-linking have been widely used to improve the properties of materials in tissue engineering. A chitosan/hydroxyapatite (CS/HA) comby scaffold with high porosity was prepared via a 3D printed pore-forming mold. The scaffold was then treated with gelatin (Gel) coating and was cross-linked by glutaraldehyde (GA) in order to improve the mechanical strength. The materials were characterized by infrared spectroscopy (IR) and X-ray diffraction (XRD). The structure of the scaffolds was observed by Scanning Electron Microscopy (SEM). Compression tests were carried out to evaluate the strength of the scaffolds. The behaviors and responses of preosteoblast cells on the scaffolds were studied as well. The results showed that gelatin coating and cross-linking significantly enhanced the mechanical strength of the porous scaffolds. Cell culture experiment indicated that the scaffold had good cytocompatibility. The combined application of 3DP structure construction and biopolymer coating/cross-linking would offer some new ideas in fabrication of porous scaffolds with enhanced strength and good biocompatibility for tissue engineering.