RESUMEN
The TFIIF-like Rpc53/Rpc37 heterodimer of RNA polymerase (pol) III is involved in various stages of transcription. The C-terminal region of Rpc53 dimerizes with Rpc37 to anchor on the lobe domain of the pol III cleft. However, structural and functional features of the Rpc53 N-terminal region had not been characterized previously. Here, we conducted site-directed alanine replacement mutagenesis on the Rpc53 N-terminus, generating yeast strains that exhibited a cold-sensitive growth defect and severely compromised pol III transcriptional activity. Circular dichroism and NMR spectroscopy revealed a highly disordered 57-amino acid polypeptide in the Rpc53 N-terminus. This polypeptide is a versatile protein-binding module displaying nanomolar-level binding affinities for Rpc37 and the Tfc4 subunit of the transcription initiation factor TFIIIC. Accordingly, we denote this Rpc53 N-terminus polypeptide as the TFIIIC-binding region or CBR. Alanine replacements in the CBR significantly reduced its binding affinity for Tfc4, highlighting its functional importance to cell growth and transcription in vitro. Our study reveals the functional basis for Rpc53's CBR in assembly of the pol III transcription initiation complex.
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ARN Polimerasa III , Factores de Transcripción TFIII , ARN Polimerasa III/metabolismo , Transcripción Genética , Factores de Transcripción TFIII/genética , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Péptidos/metabolismoRESUMEN
Aeromonas veronii is associated with food spoilage and some human diseases, such as diarrhea, gastroenteritis, hemorrhagic septicemia or asymptomatic and even death. This research investigated the mechanism of the growth, biofilm formation, virulence, stress resistance, and spoilage potential of Bacillus subtilis lipopeptide against Aeromonas veronii. Lipopeptides suppressed the transmembrane transport of Aeromonas veronii by changing the cell membrane's permeability, the structure of membrane proteins, and Na+/K+-ATPase. Lipopeptide significantly reduced the activities of succinate dehydrogenase (SDH) and malate dehydrogenase (MDH) by 86.03% and 56.12%, respectively, ultimately slowing Aeromonas veronii growth. Lipopeptides also restrained biofilm formation by inhibiting Aeromonas veronii motivation and extracellular polysaccharide secretion. Lipopeptides downregulated gene transcriptional levels related to the virulence and stress tolerance of Aeromonas veronii. Furthermore, lipopeptides treatment resulted in a considerable decrease in the extracellular protease activity of Aeromonas veronii, which restrained the decomposing of channel catfish flesh. This research provides new insights into lipopeptides for controlling Aeromonas veronii and improving food safety.
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Aeromonas , Enfermedades de los Peces , Infecciones por Bacterias Gramnegativas , Ictaluridae , Animales , Humanos , Aeromonas veronii/genética , Aeromonas veronii/metabolismo , Bacillus subtilis/genética , Biopelículas , Lipopéptidos/farmacología , Lipopéptidos/metabolismo , Infecciones por Bacterias Gramnegativas/genética , Aeromonas/genéticaRESUMEN
Recyclability of cross-link polymer materials is essential to alleviate environmental pollution caused by discarded or damaged polymers. Herein, a facile method for producing recyclable polyamide materials is developed. Linear polymer chains are constructed by Schiff base reaction between glutaraldehyde (GD) and furandiamine (FD). The linear polymer chains are crosslinked by bismaleimide (BM) to give rise to polyamide material, named GF-BMs. The resulting GF-BMs polyamide material possesses strong tensile strength (78 MPa) and good solvent resistance from room temperature to 135 °C. Especially, the thermally reversible Diels-Alder covalent bonds and dynamic imine bonds in the polymer network have a synergistic effect on fast-reprocessing, self-healing, and recyclability, which provides a new idea for recyclable materials.
RESUMEN
Food protein-derived peptides have garnered considerable attention due to their potential bioactivities and functional properties. However, the limited activity poses a challenge in effective utilization aspects. To overcome this hurdle, various methods have been explored to enhance the activity of these peptides. This comprehensive review offers an extensive overview of pretreatment, preparation methods, and modification strategies employed to augment the activity of food protein-derived peptides. Additionally, it encompasses a discussion on the current status and future prospects of bioactive peptide applications. The review also addresses the standardization of mass production processes and safety considerations for bioactive peptides while examining the future challenges and opportunities associated with these compounds. This comprehensive review serves as a valuable guide for researchers in the food industry, offering insights and recommendations to optimize the production process of bioactive peptides.
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Alimentos , Péptidos , Péptidos/químicaRESUMEN
Selective hydrogenation and hydrodeoxygenation (HDO) of biomass to value-added products play a crucial role in the development of renewable energy resources. However, achieving a temperature-controlled selectivity within one catalytic system while retaining excellent hydrogenation and HDO performance remains a great challenge. Here, nitrogen/oxygen (N/O) co-doped porous carbon nanosphere derived from resin polymer spheres is synthesized as the host matrix to in situ encapsulate highly dispersed Pd nanoparticles (NPs). Through N/O co-doping, the defects on the surface of carbon structure can serve as active sites to promote substrate adsorption. After a facile H2 O2 post-treatment process, the presence of abundant carboxyl groups on the porous carbon nanospheres can act as acidic sites to replace the use of acidic additives in the HDO process. Additionally, the increased surface oxygen-containing groups improve hydrophilicity to disperse catalysts in aqueous solutions. Owing to the unique highly dispersed Pd NPs and abundant surface defects, the Pd@APF-H2 O2 (2.3 nm) catalysts exhibit excellent catalytic activity and temperature-controlled selectivity for hydrogenation and HDO products of biomass-derived vanillin.
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Nanosferas , Biomasa , Carbono/química , Hidrogenación , Nitrógeno/química , Oxígeno/química , Porosidad , TemperaturaRESUMEN
BACKGROUND: K1 capsular polysaccharide (CPS)-associated Klebsiella pneumoniae is the primary cause of pyogenic liver abscesses (PLA) in Asia. Patients with PLA often have serious complications, ultimately leading to a mortality of ~ 5%. This K1 CPS has been reported as a promising target for development of glycoconjugate vaccines against K. pneumoniae infection. The pyruvylation and O-acetylation modifications on the K1 CPS are essential to the immune response induced by the CPS. To date, however, obtaining the fragments of K1 CPS that contain the pyruvylation and O-acetylation for generating glycoconjugate vaccines still remains a challenge. METHODS: We analyzed the digested CPS products with NMR spectroscopy and mass spectrometry to reveal a bacteriophage-derived polysaccharide depolymerase specific to K1 CPS. The biochemical and biophysical properties of the enzyme were characterized and its crystal structures containing bound CPS products were determined. We also performed site-directed mutagenesis, enzyme kinetic analysis, phage absorption and infectivity studies, and treatment of the K. pneumoniae-infected mice with the wild-type and mutant enzymes. RESULTS: We found a bacteriophage-derived polysaccharide lyase that depolymerizes the K1 CPS into fragments of 1-3 repeating trisaccharide units with the retention of the pyruvylation and O-acetylation, and thus the important antigenic determinants of intact K1 CPS. We also determined the 1.46-Å-resolution, product-bound crystal structure of the enzyme, revealing two distinct carbohydrate-binding sites in a trimeric ß-helix architecture, which provide the first direct evidence for a second, non-catalytic, carbohydrate-binding site in bacteriophage-derived polysaccharide depolymerases. We demonstrate the tight interaction between the pyruvate moiety of K1 CPS and the enzyme in this second carbohydrate-binding site to be crucial to CPS depolymerization of the enzyme as well as phage absorption and infectivity. We also demonstrate that the enzyme is capable of protecting mice from K1 K. pneumoniae infection, even against a high challenge dose. CONCLUSIONS: Our results provide insights into how the enzyme recognizes and depolymerizes the K1 CPS, and demonstrate the potential use of the protein not only as a therapeutic agent against K. pneumoniae, but also as a tool to prepare structurally-defined oligosaccharides for the generation of glycoconjugate vaccines against infections caused by this organism.
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Bacteriófagos , Infecciones por Klebsiella , Liasas , Animales , Cápsulas Bacterianas/genética , Bacteriófagos/genética , Humanos , Cinética , Klebsiella pneumoniae , RatonesRESUMEN
The rice SUB1A-1 gene, which encodes a group VII ethylene response factor (ERFVII), plays a pivotal role in rice survival under flooding stress, as well as other abiotic stresses. In Arabidopsis, five ERFVII factors play roles in regulating hypoxic responses. A characteristic feature of Arabidopsis ERFVIIs is a destabilizing N terminus, which functions as an N-degron that targets them for degradation via the oxygen-dependent N-end rule pathway of proteolysis, but permits their stabilization during hypoxia for hypoxia-responsive signaling. Despite having the canonical N-degron sequence, SUB1A-1 is not under N-end rule regulation, suggesting a distinct hypoxia signaling pathway in rice during submergence. Herein we show that two other rice ERFVIIs gene, ERF66 and ERF67, are directly transcriptionally up-regulated by SUB1A-1 under submergence. In contrast to SUB1A-1, ERF66 and ERF67 are substrates of the N-end rule pathway that are stabilized under hypoxia and may be responsible for triggering a stronger transcriptional response to promote submergence survival. In support of this, overexpression of ERF66 or ERF67 leads to activation of anaerobic survival genes and enhanced submergence tolerance. Furthermore, by using structural and protein-interaction analyses, we show that the C terminus of SUB1A-1 prevents its degradation via the N-end rule and directly interacts with the SUB1A-1 N terminus, which may explain the enhanced stability of SUB1A-1 despite bearing an N-degron sequence. In summary, our results suggest that SUB1A-1, ERF66, and ERF67 form a regulatory cascade involving transcriptional and N-end rule control, which allows rice to distinguish flooding from other SUB1A-1-regulated stresses.
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Proteínas de Arabidopsis/genética , Proteínas de Unión al ADN/genética , Oryza/genética , Proteínas de Plantas/genética , Estrés Fisiológico/genética , Factores de Transcripción/genética , Adaptación Fisiológica/genética , Anaerobiosis/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/genética , Oryza/crecimiento & desarrollo , Transducción de Señal/genética , Especificidad por SustratoRESUMEN
Vibrio cholerae is the causative agent of the diarrheal disease cholera, for which biofilm communities are considered to be environmental reservoirs. In endemic regions, and after algal blooms, which may result from phosphate enrichment following agricultural runoff, the bacterium is released from biofilms resulting in seasonal disease outbreaks. However, the molecular mechanism by which V. cholerae senses its environment and switches lifestyles from the biofilm-bound state to the planktonic state is largely unknown. Here, we report that the major biofilm scaffolding protein RbmA undergoes autocatalytic proteolysis via a phosphate-dependent induced proximity activation mechanism. Furthermore, we show that RbmA mutants that are defective in autoproteolysis cause V. cholerae biofilms to grow larger and mechanically stronger, correlating well with the observation that RbmA stability directly affects microbial community homeostasis and rheological properties. In conclusion, our biophysical study characterizes a novel phosphate-dependent breakdown pathway of RbmA, while microbiological data suggest a new, sensory role of this biofilm scaffolding element.
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Proteínas Bacterianas/metabolismo , Biopelículas/efectos de los fármacos , Compuestos de Magnesio/farmacología , Fosfatos/farmacología , Proteolisis , Vibrio cholerae/metabolismo , Proteínas Bacterianas/genética , Biopelículas/crecimiento & desarrollo , Vibrio cholerae/efectos de los fármacos , Vibrio cholerae/crecimiento & desarrolloRESUMEN
Recent research on the structure and mechanism of DNA polymerases has continued to generate fundamentally important features, including a noncanonical pathway involving "prebinding" of metal-bound dNTP (MdNTP) in the absence of DNA. While this noncanonical mechanism was shown to be a possible subset for African swine fever DNA polymerase X (Pol X) and human Pol λ, it remains unknown whether it could be the primary pathway for a DNA polymerase. Pol µ is a unique member of the X-family with multiple functions and with unusual Mn2+ preference. Here we report that Pol µ not only prebinds MdNTP in a catalytically active conformation but also exerts a Mn2+ over Mg2+ preference at this early stage of catalysis, for various functions: incorporation of dNTP into a single nucleotide gapped DNA, incorporation of rNTP in the nonhomologous end joining (NHEJ) repair, incorporation of dNTP to an ssDNA, and incorporation of an 8-oxo-dGTP opposite template dA (mismatched) or dC (matched). The structural basis of this noncanonical mechanism and Mn2+ over Mg2+ preference in these functions was analyzed by solving 19 structures of prebinding binary complexes, precatalytic ternary complexes, and product complexes. The results suggest that the noncanonical pathway is functionally relevant for the multiple functions of Pol µ. Overall, this work provides the structural and mechanistic basis for the long-standing puzzle in the Mn2+ preference of Pol µ and expands the landscape of the possible mechanisms of DNA polymerases to include both mechanistic pathways.
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ADN Polimerasa Dirigida por ADN/metabolismo , Manganeso/metabolismo , ADN Polimerasa Dirigida por ADN/química , ADN Polimerasa Dirigida por ADN/aislamiento & purificación , Humanos , Manganeso/química , Modelos MolecularesRESUMEN
The vast majority of in vitro structural and functional studies of the activation mechanism of protein kinases use the kinase domain alone. Well-demonstrated effects of regulatory domains or allosteric factors are scarce for serine/threonine kinases. Here we use a site-specifically phosphorylated SCD1-FHA1-kinase three-domain construct of the serine/threonine kinase Rad53 to show the effect of phospho-priming, an in vivo regulatory mechanism, on the autophosphorylation intermediate and specificity. Unphosphorylated Rad53 is a flexible monomer in solution but is captured in an asymmetric enzyme:substrate complex in crystal with the two FHA domains separated from each other. Phospho-priming induces formation of a stable dimer via intermolecular pT-FHA binding in solution. Importantly, autophosphorylation of unprimed and phospho-primed Rad53 produced predominantly inactive pS350-Rad53 and active pT354-Rad53, respectively. The latter mechanism was also demonstrated in vivo. Our results show that, while Rad53 can display active conformations under various conditions, simulation of in vivo regulatory conditions confers functionally relevant autophosphorylation.
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Proteínas de Ciclo Celular/química , Proteínas de Ciclo Celular/metabolismo , Quinasa de Punto de Control 2/química , Quinasa de Punto de Control 2/metabolismo , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas de Ciclo Celular/genética , Quinasa de Punto de Control 2/genética , Daño del ADN , Modelos Moleculares , Resonancia Magnética Nuclear Biomolecular , Fosforilación , Fosfotreonina/metabolismo , Dominios Proteicos , Multimerización de Proteína , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Dispersión del Ángulo Pequeño , Serina/química , Treonina/química , Treonina/metabolismoRESUMEN
The mechanism of DNA polymerase (pol) fidelity is of fundamental importance in chemistry and biology. While high-fidelity pols have been well studied, much less is known about how some pols achieve medium or low fidelity with functional importance. Here we examine how human DNA polymerase λ (Pol λ) achieves medium fidelity by determining 12 crystal structures and performing pre-steady-state kinetic analyses. We showed that apo-Pol λ exists in the closed conformation, unprecedentedly with a preformed MgdNTP binding pocket, and binds MgdNTP readily in the active conformation in the absence of DNA. Since prebinding of MgdNTP could lead to very low fidelity as shown previously, it is attenuated in Pol λ by a hydrophobic core including Leu431, Ile492, and the Tyr505/Phe506 motif. We then predicted and demonstrated that L431A mutation enhances MgdNTP prebinding and lowers the fidelity. We also hypothesized that the MgdNTP-prebinding ability could stabilize a mismatched ternary complex and destabilize a matched ternary complex, and provided evidence with structures in both forms. Our results demonstrate that, while high-fidelity pols follow a common paradigm, Pol λ has developed specific conformations and mechanisms for its medium fidelity. Structural comparison with other pols also suggests that different pols likely utilize different conformational changes and microscopic mechanisms to achieve their catalytic functions with varying fidelities.
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ADN Polimerasa beta/química , ADN Polimerasa beta/metabolismo , Cristalografía por Rayos X , ADN Polimerasa beta/genética , Humanos , Modelos Moleculares , Conformación ProteicaRESUMEN
Presystemic interactions with gut microbiota might play important roles in the holistic action of herbal medicines in their traditional oral applications. However, research interests usually focus on biologic activities of the in vivo available herb-derived components and their exposure in circulation. In this study, we illustrated the importance of studying the presystemic interplay with gut microbiota for understanding the holistic actions of medicinal herbs by using calycosin-7-O-ß-D-glucoside (C7G), the most abundant flavonoid and chemical marker in Astragali Radix, as a model compound. When C7G was orally administrated to rats, calycosin-3'-O-glucuronide (G2) was the major circulating component in the blood together with a minor calycosin but not C7G. Rat gut microbiota hydrolyzed C7G in vitro rapidly and produced its aglycone calycosin. Calycosin exhibited higher permeability than C7G and further underwent extensive glucuronidation to yield 3'-glucuronide as the dominant metabolite. Bioactivity assays revealed that G2 exhibited similar or more potent proangiogenic effects than calycosin in human umbilical vein endothelial cells in vitro and in the vascular endothelial growth factor receptor tyrosine kinase inhibitor II-induced blood vessel loss model in zebrafish. More interestingly, the incubation of C7G with gut microbiota from both normal and colitic rats showed a probiotics-like effect through stimulating the growth of the beneficial bacteria Lactobacillus and Bifidobacterium. In conclusion, C7G interacts reciprocally with gut microbiota after oral dosing, which makes it not only an angiogenic prodrug but also a modulator of gut microbiota.
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Microbioma Gastrointestinal/efectos de los fármacos , Microbioma Gastrointestinal/fisiología , Glucósidos/administración & dosificación , Glucósidos/metabolismo , Isoflavonas/administración & dosificación , Isoflavonas/metabolismo , Administración Oral , Animales , Células CACO-2 , Colitis/metabolismo , Humanos , Masculino , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
A dogma for DNA polymerase catalysis is that the enzyme binds DNA first, followed by MgdNTP. This mechanism contributes to the selection of correct dNTP by Watson-Crick base pairing, but it cannot explain how low-fidelity DNA polymerases overcome Watson-Crick base pairing to catalyze non-Watson-Crick dNTP incorporation. DNA polymerase X from the deadly African swine fever virus (Pol X) is a half-sized repair polymerase that catalyzes efficient dG:dGTP incorporation in addition to correct repair. Here we report the use of solution structures of Pol X in the free, binary (Pol X:MgdGTP), and ternary (Pol X:DNA:MgdGTP with dG:dGTP non-Watson-Crick pairing) forms, along with functional analyses, to show that Pol X uses multiple unprecedented strategies to achieve the mutagenic dG:dGTP incorporation. Unlike high fidelity polymerases, Pol X can prebind purine MgdNTP tightly and undergo a specific conformational change in the absence of DNA. The prebound MgdGTP assumes an unusual syn conformation stabilized by partial ring stacking with His115. Upon binding of a gapped DNA, also with a unique mechanism involving primarily helix αE, the prebound syn-dGTP forms a Hoogsteen base pair with the template anti-dG. Interestingly, while Pol X prebinds MgdCTP weakly, the correct dG:dCTP ternary complex is readily formed in the presence of DNA. H115A mutation disrupted MgdGTP binding and dG:dGTP ternary complex formation but not dG:dCTP ternary complex formation. The results demonstrate the first solution structural view of DNA polymerase catalysis, a unique DNA binding mode, and a novel mechanism for non-Watson-Crick incorporation by a low-fidelity DNA polymerase.
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Virus de la Fiebre Porcina Africana/enzimología , ADN Polimerasa Dirigida por ADN/química , ADN Polimerasa Dirigida por ADN/metabolismo , ADN/metabolismo , Fiebre Porcina Africana/virología , Virus de la Fiebre Porcina Africana/química , Virus de la Fiebre Porcina Africana/metabolismo , Animales , Emparejamiento Base , ADN/química , ADN Polimerasa beta/química , ADN Polimerasa beta/metabolismo , Nucleótidos de Desoxicitosina/metabolismo , Nucleótidos de Desoxiguanina/metabolismo , Guanosina Trifosfato/metabolismo , Modelos Moleculares , Resonancia Magnética Nuclear Biomolecular , Conformación Proteica , Porcinos/virologíaRESUMEN
A sensitive and specific LC-MS/MS method was developed for simultaneous determination of aloe-emodin, rhein, emodin, chrysophanol and physcion and their conjugates in rat plasma. The lower limit of quantitation of each anthraquinone was 0.020-0.040 µm. Intra-day and inter-day accuracies were 90.1-114.3% and the precisions were <14.6%. The matrix effects were 104.0-113.2%. The method was successfully applied to a pharmacokinetic study in rats receiving a rhubarb extract orally. The area under the concentration-time curve (AUC0-t ) and peak concentration (Cmax ) of free aloe-emodin and emodin in rat plasma were much lower than those of rhein. The amounts of chrysophanol and physcion were too low to be continuously detected. After treating the plasma samples with ß-glucuronidases, each anthraquinone was detectable throughout the experimental period (36 h) and showed much higher plasma concentrations and AUC0-t . The free/total ratios of aloe-emodin, rhein and emodin were 6.5, 49.0 and 1.7% for Cmax and 3.7, 32.5 and 1.1% for AUC0-t , respectively. The dose-normalized AUC0-t and Cmax of the total of each anthraquinone were in the same descending order: rhein > emodin > chrysophanol > physcion > aloe-emodin. These findings reveal phase II conjugates as the dominant in vivo existing forms of rhubarb antharquinones and warrant a further study to evaluate their contribution to the herbal activity.
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Antraquinonas/sangre , Antraquinonas/farmacocinética , Extractos Vegetales/administración & dosificación , Rheum/química , Animales , Antraquinonas/química , Masculino , Extractos Vegetales/química , Ratas , Ratas Sprague-DawleyRESUMEN
PURPOSE: To investigate the impact of rapamycin on the differentiation of hair cells. METHODS: Murine cochlear organoids were derived from cochlear progenitor cells. Different concentrations of rapamycin were added into the culture medium at different proliferation and differentiation stages. RESULTS: Rapamycin exhibited a concentration-dependent reduction in the proliferation of these inner ear organoids. Nevertheless, organoids subjected to a 10-nM dose of rapamycin demonstrated a markedly increased proportion of hair cells. Furthermore, rapamycin significantly upregulated the expression of markers associated with both hair cells and supporting cells, including ATOH1, MYO7A, and SOX2. Mechanistic studies revealed that rapamycin preferentially suppressed cells without Sox2 expression during the initial proliferation stage, thereby augmenting and refining the population of SOX2+ progenitors. These enriched progenitors were predisposed to differentiate into hair cells during the later stages of organoid development. Conversely, the use of the mTOR activator MHY 1485 demonstrated opposing effects. CONCLUSION: Our findings underscore a practical strategy for enhancing the generation of inner ear organoids with a low dose of rapamycin, achieved by enriching SOX2+ progenitors in an in vitro setting.
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Oído Interno , Sirolimus , Animales , Ratones , Animales Recién Nacidos , Diferenciación Celular/efectos de los fármacos , Oído Interno/efectos de los fármacos , Organoides/efectos de los fármacos , Sirolimus/farmacología , Factores de Transcripción SOXB1/metabolismoRESUMEN
BACKGROUND: Our previous research demonstrated that resveratrol counters DDP-induced ototoxicity by upregulating miR-455-5p, which targets PTEN. This study aimed to elucidate the underlying mechanisms involving GAS5 and DNA methyltransferase 1 (DNMT1) in resveratrol's protective action. METHODS: A luciferase reporter assay and RNA immunoprecipitation (RIP) assay were employed to study the binding between GAS5 and miR-455-5p, as well as between miR-455-5p and PTEN. HEI-OC1 cells treated with DDP were transfected with vectors for GAS5, si-GAS5, DNMT1, si-DNMT1, and miR-455-5p mimics, as well as PTEN. Subsequently, they were treated with resveratrol and exposed to DDP, both separately and in combination. The distribution of CpG islands in the GAS5 promoter was identified using MethyPrimer, and methylation-specific PCR (MSP) was conducted to determine the methylation levels of GAS5. Chromatin immunoprecipitation (ChIP) was utilized to examine the interaction between DNMT1 and GAS5. The viability of HEI-OC1 cells, catalase (CAT) activity, apoptosis, and ROS levels were assessed using the CCK-8 assay, CAT assay, TUNEL staining, and flow cytometry, respectively. An in vivo mouse model was developed to measure auditory brainstem response (ABR) thresholds, while RT-qPCR and Western blot analysis were employed to evaluate molecular levels. RESULTS: Our study discovered that GAS5 acts as a sponge for miR-455-5p, thereby increasing PTEN expression in DDP-treated HEI-OC1 cells. This process was reversed upon treatment with resveratrol. Importantly, DNMT1 promoted the methylation of the GAS5 promoter, leading to the suppression of GAS5 expression. This suppression enhanced the effectiveness of resveratrol in combating DDP-induced apoptosis and ROS in HEI-OC1 cells and amplified its protective effect against DDP's ototoxicity in vivo. CONCLUSIONS: Our research emphasizes the significance of the DNMT1/GAS5/miR-455-5p/PTEN axis as a promising new route to boost resveratrol's effectiveness against DDP-induced ototoxicity.
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Cisplatino , ADN (Citosina-5-)-Metiltransferasa 1 , Epigénesis Genética , MicroARNs , Ototoxicidad , Fosfohidrolasa PTEN , ARN Largo no Codificante , Resveratrol , Fosfohidrolasa PTEN/metabolismo , Fosfohidrolasa PTEN/genética , MicroARNs/genética , MicroARNs/metabolismo , Animales , Resveratrol/farmacología , Resveratrol/uso terapéutico , Ratones , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Ototoxicidad/prevención & control , ADN (Citosina-5-)-Metiltransferasa 1/metabolismo , ADN (Citosina-5-)-Metiltransferasa 1/genética , Epigénesis Genética/efectos de los fármacos , Línea Celular , Masculino , Ratones Endogámicos C57BL , Transducción de Señal/efectos de los fármacos , Apoptosis/efectos de los fármacosRESUMEN
Adjustments in foliar photoprotective pigments are crucial for plant adaptation to harsh environments, serving as indicators of environmental stress. However, understanding when and where these adjustments occur across diverse biomes remains unclear due to challenges in large-scale observation. Here, we propose a novel approach to assess dynamics in photoprotective pigments at the canopy level using a new index derived from space-borne optical sensors. This approach generates a global map depicting the daily mean shortwave radiation threshold at which adjustments typically occur under prevailing climatic conditions. The global average of this threshold is 262 ± 50 W mâ»2, with lower values at high latitudes and peaks near 40° in both hemispheres. Temperature exerts a stronger influence on this latitudinal pattern than humidity. Future projections suggest a decrease in this threshold over northern high latitudes, implying exacerbated vulnerability under identical radiation levels due to negative warming responses. Based on this threshold, a high-stress zone around 60°N is identified and is predicted to shift southward in the future. These findings bridge critical gaps in photoprotection research and offer a new perspective on understanding the biogeochemical cycles of global ecosystems. This framework can also enhance our ability to predict the fate of diverse ecosystems under future climate.
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The theories for substrate recognition in enzyme catalysis have evolved from lock-key to induced fit, then conformational selection, and conformational selection followed by induced fit. However, the prevalence and consensus of these theories require further examination. Here we use cryogenic electron microscopy and African swine fever virus type 2 topoisomerase (AsfvTop2) to demonstrate substrate binding theories in a joint and ordered manner: catalytic selection by the enzyme, conformational selection by the substrates, then induced fit. The apo-AsfvTop2 pre-exists in six conformers that comply with the two-gate mechanism directing DNA passage and release in the Top2 catalytic cycle. The structures of AsfvTop2-DNA-inhibitor complexes show that substantial induced-fit changes occur locally from the closed apo-conformer that however is too far-fetched for the open apo-conformer. Furthermore, the ATPase domain of AsfvTop2 in the MgAMP-PNP-bound crystal structures coexist in reduced and oxidized forms involving a disulfide bond, which can regulate the AsfvTop2 function.
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Lighting conditions are an important factor affecting dry-cured products. This study investigated the effects of treatments with different light intensities (0 lx, 1000 lx, 25000 lx) and different light sources including red light, blue light, UV-light on oxidation leve and flavor change in dry-cured Wuchang fish. The results showed that dry-cured Wuchang fish exhibited an attractive brown-yellow color, the highest oxidation degree of myoglobin (Mb), the highest fat oxidation under the light conditions of 25000 lx light intensity and UV-light irradiation. This phenomenon was observed that the degree of Mb oxidation was increased, while the degree of fat oxidation was increased. At 25000 lx light intensity and UV-light irradiation, dry-cured Wuchang fish showed an ignificantly decreased fatty acid conten (especially oleic acid and linoleic acid), significantly increased characteristic volatile compound contents (22 for 25,000 lx light intensity and 27 for UV-light irradiation), which contributed to the improvement of quality stability of dry-cured Wuchang fish. Our findings provide theoretical support for the industrial application of exogenous light in dry-cured Wuchang fish.