Zanthoxylum bungeanum is an economically important crop worldwide due to its high content of aroma-producing monoterpenoids,and development of varieties with enhanced flavor and overall quality is a crucial research a...Zanthoxylum bungeanum is an economically important crop worldwide due to its high content of aroma-producing monoterpenoids,and development of varieties with enhanced flavor and overall quality is a crucial research area.However,the transcriptional regulatory mechanisms underlying monoterpenoid synthesis in Z.bungeanum remain unclear,hindering these breeding efforts.In this study,RNA sequencing,gas chromatography–mass spectrometry,and other molecular biology techniques were used to identify the underlying transcriptional regulation mechanisms.Two transcription factors,ZbbHLH2 and ZbERF6,were identified as key regulators of monoterpenoid synthesis in Z.bungeanum that upregulate various monoterpenoid synthesis-associated genes and are novel transcriptional activators of ZbIDI,which encodes the rate-limiting enzyme in plant monoterpenoid synthesis.Functional analysis revealed that the expression of three genes[1]modulates monoterpenoid accumulation in Z.bungeanum peel.These findings provide novel insights into the metabolic regulatory network of monoterpenoid synthesis in Z.bungeanum peel,offer potential strategies for the biofortification of specific monoterpenoids,and will promote the development of Z.bungeanum germplasm for targeted breeding and quality improvement.展开更多
Anthocyanins are important flavonoid pigments in the coloration of fruits.To identify candidate genes involved in anthocyanin accumulation,metabolic and transcriptome analyses were conducted in‘Nanguo'pear and it...Anthocyanins are important flavonoid pigments in the coloration of fruits.To identify candidate genes involved in anthocyanin accumulation,metabolic and transcriptome analyses were conducted in‘Nanguo'pear and its red sport cultivar‘Nanhong'pear.The results showed that‘Nanhong'pear had significantly higher anthocyanin and flavonol contents.Additionally,transcriptomic analysis showed that there were significant differences in the expression of genes involved in phenylpropanoid and flavonoid biosynthesis pathways between the two cultivars,with PuGSTF12 being the most upregulated gene in the‘Nanhong'cultivar.Further analysis identified a novel MYB transcription factor(PuMYB93),and its silencing repressed PuGSTF12 expression and anthocyanin accumulation,suggesting it plays an essential role in the regulation of anthocyanin biosynthesis.Moreover,yeast one-hybrid analysis,electrophoretic mobility shift assay,andβ-glucuronidase assay indicated that PuMYB93 can directly bind to the PuGSTF12 promoter to positively regulate its expression.Additionally,PuGSTF12 silencing suppressed the coloration of PuMYB93-OE peels,suggesting that PuGSTF12 act downstream of PuMYB93.Overall,the findings of this study show that PuMYB93 promotes anthocyanin transport in pears by regulating PuGSTF12 expression to further enhance anthocyanin accumulation.展开更多
Objectives:Oxidative stress(OS)plays a pivotal role in chronic and neurodegenerative diseases,which has sparked interest in molecules that modulate redox-regulating enzymes.Melatonin and its metabolites exhibit antiox...Objectives:Oxidative stress(OS)plays a pivotal role in chronic and neurodegenerative diseases,which has sparked interest in molecules that modulate redox-regulating enzymes.Melatonin and its metabolites exhibit antioxidant properties;however,their molecular mechanisms of enzymatic and transcriptional modulation remain unclear.This study aimed to investigate,through an exploratory in silico approach,the interactions of melatonin and related compounds with OS-related enzymes to generate hypotheses about their role in cellular redox control.Methods:A rational selection of antioxidant,pro-oxidant,and transcriptional targets was performed.Ligands were optimized at the DFT level(M05-2X/6-311+G(d,p))and docked to OS related enzymes.Docking results were analyzed using polygenic antioxidant indices(PAOX)and a similarity interaction index(SSI).Molecular dynamics simulations of selected complexes provided additional insight into potential ligand-protein interaction mechanisms.Results:In silico analyses revealed that N1-acetyl-5-methoxykynuramine(AMK),N1-acetyl-N2-formyl-5-methoxykynuramine(AFMK),and 3-hydroxymelatonin(3OH-M)could partially inhibit pro-oxidant enzymes such as neuronal nitric oxide synthase(nNOS),5-lipoxygenase(5-LOX),thioredoxin reductase(TrxR),and nicotinamide adenine dinucleotide phosphate oxidase(NOX5).The N-(2-(2-acetyl-6,7-dihydroxy-1H-indol-3-yl)ethyl)acetamide(IIcD)and N-(2-(6-hydroxy-7-mercapto-5-methoxy-1H-indol)ethyl)acetamide(dM38)derivatives could potentially stabilize superoxide dismutase(SOD1)and catalase(CAT)enzymes,respectively.Finally,AFMK and dM38 showed consistent interactions with transcriptional regulators,particularly peroxisome proliferator-activated receptor alpha(PPARα)and Kelchlike ECH-associated protein 1(KEAP1).Conclusion:These studies about melatonin-related compounds support a multifactorial profile of redox modulation and provide mechanistic hypotheses for future experimental validation.Among these approaches,the interaction-similarity index is introduced as a novel tool to facilitate the identification of promising redox-active candidates.展开更多
Flavonoids,abundant in the fruits,are pivotal to their growth,development,and storage.In addition,they have significant beneficial effects on human health.Consequently,research is increasingly concentrating on the reg...Flavonoids,abundant in the fruits,are pivotal to their growth,development,and storage.In addition,they have significant beneficial effects on human health.Consequently,research is increasingly concentrating on the regulatory mechanisms governing flavonoid biosynthesis in fruits.Phytohormones are involved in the regulation of flavonoid biosynthesis.The abscisic acid,ethylene,jasmonic acid,cytokinins,and brassinosteroids promote flavonoid biosynthesis,while auxin negatively regulates flavonoid biosynthesis.Subsequently,transcription factors from the MYB,bHLH,WRKY,NAC,and bZIP families are pivotal in regulating flavonoid biosynthesis.In addition,non-coding RNAs(microRNA and lncRNA)also participate in the regulation of flavonoids biosynthesis.MicroRNAs are generally believed to negatively regulate flavonoid metabolism in fruits,while lncRNAs have the opposite effect.Furthermore,the interactions between plant hormones,transcription factors,and non-coding RNAs in fruit flavonoid biosynthesis were analyzed.Ultimately,a foundational regulatory network for fruit flavonoid biosynthesis was hereby established.展开更多
Anthocyanins are the main pigments in ripe strawberry fruits.FaMYB10 and abscisic acid(ABA)are the two major regulators of anthocyanin biosynthesis in the maturation process of strawberry fruits.However,the transcript...Anthocyanins are the main pigments in ripe strawberry fruits.FaMYB10 and abscisic acid(ABA)are the two major regulators of anthocyanin biosynthesis in the maturation process of strawberry fruits.However,the transcriptional regulation of FaMYB10,ABA biosynthesis,and anthocyanin accumulation in strawberry(Fragaria×ananassa)remain largely unknown.Here,a yeast one-hybrid screen using the FaMYB10 promoter identified a class B MADS-domain transcription factor,FaMADS6 in strawberry.FaMADS6 exhibited high expression at the early developmental stage but was hardly detected during maturation of strawberry fruit,a pattern opposite to accumulation of anthocyanins.Transcriptional upregulation of FaMADS6 repressed anthocyanin accumulation and expression of FaMYB10 and the anthocyanin biosynthetic genes,FaCHS,FaCHI,FaF3H,FaANS,and FaUFGT.In contrast,downregulation of FaMADS6 promoted the expression of FaMYB10 and the anthocyanin biosynthetic genes.The promoters of the anthocyanin biosynthetic genes were not directly bound by FaMADS6,in contrast to FaMYB10.Analysis of the DNA binding sequences of FaMADS6 revealed that it also interacted with the promoters of FaNCED2 and FaPYR1,which are involved in the biosynthesis and perception of ABA.Overexpression of FaMADS6 significantly suppressed FaNCED2 and FaPYR1 and ABA synthesis in transgenic strawberry.Together,our findings suggest that FaMADS6 functions as a suppressor of anthocyanin accumulation by directly downregulating FaMYB10 and ABA production during strawberry fruit maturation.展开更多
Tobacco(Nicotiana tabacum,2n=48)is a key non-food economic crop,yet its stress response and gene regulatory mechanisms remain poorly understood.By analyzing 603 transcriptome datasets,this study identified 1405 tissue...Tobacco(Nicotiana tabacum,2n=48)is a key non-food economic crop,yet its stress response and gene regulatory mechanisms remain poorly understood.By analyzing 603 transcriptome datasets,this study identified 1405 tissue-specific genes,revealing tissue-specific synthesis of terpenoids and other ecologically important secondary metabolites in sepals and other tissues.Comparative stress-response analysis highlighted distinct gene expression patterns in leaves and roots under biotic and abiotic stresses.Additionally,28,396 expression quantitative trait loci(eQTLs)were mapped in leaves,offering valuable genetic regulatory markers.These findings provide crucial insights into tobacco’s gene expression characteristics and their functional implications,serving as a foundation for future research.展开更多
Flavonoids are crucial secondary metabolites widely distributed in plants,playing vital roles in diverse biological processes.Although the flavonoid biosynthesis pathway has been extensively characterized,the transcri...Flavonoids are crucial secondary metabolites widely distributed in plants,playing vital roles in diverse biological processes.Although the flavonoid biosynthesis pathway has been extensively characterized,the transcriptional regulatory mechanisms remain poorly understood.In this study,we identify the miR166–ATHB14-LIKE module comprising the miR166 and its target gene ATHB14-LIKE as a key regulator of flavonoid biosynthesis in soybean(Glycine max).Knockdown of miR166 or overexpression of ATHB14-LIKE upregulated multiple flavonoid biosynthesis genes,leading to increased flavonoid accumulation.Conversely,miR166 overexpression suppressed these genes and reduced flavonoid levels.We further show that ATHB14-LIKE directly activates specific flavonoid biosynthesis genes by binding to their promoters.Additionally,ATHB14-LIKE forms homodimers and heterodimers with homologous proteins to regulate downstream flavonoid biosynthesis genes.Together,our findings demonstrate that the miR166–ATHB14-LIKE module controls soybean flavonoid content by coordinating the expression of key biosynthetic genes.展开更多
Drought stress significantly impedes apple growth,development,and yield,leading to substantial economic losses within the global apple industry.Malus prunifolia(Mp),a commonly utilized apple rootstock,has shown promis...Drought stress significantly impedes apple growth,development,and yield,leading to substantial economic losses within the global apple industry.Malus prunifolia(Mp),a commonly utilized apple rootstock,has shown promise in augmenting cultivated apple resistance to abiotic stress.Although Alfin-like(ALs)proteins have demonstrated pivotal roles in dicotyledonous plants'response to abiotic stresses,knowledge about AL genes in apple rootstocks is limited,and their functions remain largely elusive.In this study,we identified and characterized 10 MpAL gene members in the apple rootstock genome,confirming their localization within the nucleus.Our investigation revealed the significant regulation of MpALs'expression under drought and abscisic acid(ABA)stresses in M.prunifolia.In this study,one of the members,MpAL1,was selected for further exploration in Arabidopsis and apple to explore its potential function in response to drought and ABA stresses.The results showed that overexpression-MpAL1 transgenic apple calli grew significantly better than WT and MpAL1-RNAi lines,which regulates the accumulation of H_(2)O_(2)and O_(2).-levels.Additionally,transgenic Arabidopsis plants overexpressing MpAL1 exhibited positively regulating antioxidant enzymes activities under stress treatments.Further study showed that silencing MpAL1 in apple plants showed obvious chlorosis in leaves,and accumulation of reactive oxygen species under drought stress.Moreover,our detailed analysis established that MpAL1 regulates several drought and ABA-responsive genes,exerting an influence on their expression in transgenic apple.Collectively,our findings identify MpAL1 as a positive regulator that increases drought stress in apple,shedding light on its potential significance in bolstering drought resistance in this fruit crop.展开更多
Anthocyanins play a crucial role in shaping the visual appeal and nutritional quality of fruits.Previous research on anthocyanin biosynthesis in sweet cherry(Prunus avium L.)has primarily relied on single-omics approa...Anthocyanins play a crucial role in shaping the visual appeal and nutritional quality of fruits.Previous research on anthocyanin biosynthesis in sweet cherry(Prunus avium L.)has primarily relied on single-omics approaches or focused on a limited range of metabolites,leaving the regulatory mechanisms and dynamic metabolism of anthocyanins during ripening inadequately characterized.This study integrated anthocyanin-targeted metabolomics and transcriptomics to identify key anthocyanins in sweet cherry and construct a transcriptional regulatory network for anthocyanin biosynthesis.A novel bHLH transcription factor,Prunus avium bHLH transcription factor 102(PavbHLH102),was identified,and its role in regulating cyanidin levels was validated through overexpression and silencing experiments.Both in vitro and in vivo assays demonstrated that PavbHLH102 activates key anthocyanin biosynthetic genes,including PavF3H,PavDFR,and PavUFGT,thereby enhancing fruit coloration.Notably,PavF3′H upregulation significantly increased cyanidin accumulation.This study provides new insights into anthocyanin regulation in sweet cherry and offers valuable resources for improving fruit quality.展开更多
Background:Aberrant expression of transcription factors(TFs)is a key mechanism mediating tumor immune escape and therapeutic resistance.The involvement of E26 transformation-specific(ETS)family of TFs in immune regula...Background:Aberrant expression of transcription factors(TFs)is a key mechanism mediating tumor immune escape and therapeutic resistance.The involvement of E26 transformation-specific(ETS)family of TFs in immune regulation is not fully understood.The study aimed to elucidate the function of E-twenty-six variant 4(ETV4)in tumor immune evasion and its potential as a predictive biomarker for immunotherapy in melanoma.Methods:The expression patterns of ETS family TFs were analyzed in melanoma and hepatocellular carcinoma(HCC).Single-cell RNA sequencing(scRNA-seq)was used to dissect the cellular expression and function of ETV4 in the tumor microenvironment.Functional studies and murine models were employed to investigate the role of ETV4 in T cell-mediated tumor killing and tumor growth.The correlation between ETV4 expression level and patient responsiveness to programmed cell death protein 1(PD-1)blockade therapy was evaluated.Results:TFs in the ETS family were found to effectively stratify melanoma and HCC patients into prognostic subgroups.In melanoma,the polyoma enhancer activator 3(PEA3)subfamily,particularly ETV4 and ETV5,showed a negative correlation with immune infiltration.scRNA-seq analysis showed that ETV4 is preferentially expressed in melanoma cells and involves in mediating tumor-immunocyte interactions.Functional studies demonstrated that ETV4 impairs T cell-mediated tumor killing by transcriptionally upregulating programmed death-ligand 1(PD-L1).In immunocompetent murine models,ETV4 downregulation significantly suppressed tumor growth.Furthermore,high ETV4 expression correlated with poor responses to anti-PD-1 therapy.Conclusion:Our findings identify ETV4 as a key transcriptional regulator of immune evasion in melanoma by controlling PD-L1 expression.ETV4 may act as a predictive biomarker for immunotherapy outcomes.展开更多
Flower color is an essential trait in ornamental plant breeding. Lycoris longituba is a popular ornamental plant native to central eastern China. The decrease in anthocyanin accumulation causes L. longituba petal colo...Flower color is an essential trait in ornamental plant breeding. Lycoris longituba is a popular ornamental plant native to central eastern China. The decrease in anthocyanin accumulation causes L. longituba petal color fading during flower development, which considerably affects the ornamental value of L. longituba. However, mechanisms underlying anthocyanin biosynthesis inhibition during L. longituba petal development remain unclear. In this study, three LlDFR genes were confirmed to be involved in anthocyanin biosynthesis and LlDFRc exerted the strongest promoting effect on anthocyanin accumulation. According to the correlation analysis results, LlbHLH12 exhibited the strongest negative correlation with LlDFRc. Quantitative real-time PCR analysis showed that LlbHLH12 was highly expressed during the medium bud and full bloom stages of flower development. LlbHLH12 was identified as a member of subgroup XII of bHLH transcription factor family. Subcellular localization and transcriptional activation ability assay revealed that LlbHLH12 was located in the nucleus without transcriptional activation activity. Overexpression of LlbHLH12 in Nicotiana tabacum and L. longituba inhibited anthocyanin accumulation by suppressing the expression of anthocyanin biosynthetic pathway genes. Furthermore, yeast one-hybrid, dual-luciferase, and β-glucuronidase activity assays showed that LlbHLH12 directly bound to the promoters of LlPAL and LlDFRc and suppressed their expression to inhibit anthocyanin biosynthesis. Overall, our study identified a novel bHLH repressor negatively regulating anthocyanin biosynthesis and provided new insights into the molecular mechanisms underlying color fading in L. longituba petals.展开更多
The B-box(BBX)gene family plays a vital role in plant growth,development,and stress responses.This study aimed to characterize the SmBBX gene family in eggplant(Solanum melongena L.),addressing the lack of systematic ...The B-box(BBX)gene family plays a vital role in plant growth,development,and stress responses.This study aimed to characterize the SmBBX gene family in eggplant(Solanum melongena L.),addressing the lack of systematic bioinformatics and functional studies in this species.A total of 33 SmBBX genes were identified through genome-wide analysis.These genes were phylogenetically grouped into five major clades,with shared domain structures,motifs,and genomic architectures among clade members.The gene duplication analysis revealed segmental duplication as the primary mechanism underlying the expansion of SmBBX proteins in eggplant.Additionally,expression profiling across diverse tissues and abiotic stress conditions,combined with the construction of protein—protein interaction networks and luciferase complementation assay,provided valuable insights into the functional roles of SmBBX genes.SmBBX21-2 and SmBBX22 were identified as the key regulators of anthocyanin biosynthesis,activating the expression of SmCHS and SmDFR promoters.Functional validation via heterologous and homologous overexpression demonstrated that SmBBX22 promoted anthocyanin accumulation by upregulating the expression of structural genes(SmCHS,SmF3H,SmF3′5′H,SmDFR,and SmANS)and transcription factors(SmTT8 and SmHY5)important for anthocyanin biosynthesis.Furthermore,the integration of DNA affinity purification sequencing and RNA-seq data revealed the direct transcriptional targets of SmBBX22,including genes involved in secondary metabolism,hormone signaling,and developmental regulation.This highlighted the role of SmBBX22 in phenylpropanoid and flavonoid biosynthesis.This study lays the foundation for understanding the functional roles of BBX genes in eggplant and provides new directions for future research in plant metabolism and stress adaptation.展开更多
Background:The regulatory mechanisms governing vasculogenic mimicry(VM)in oral squamous cell carcinoma(OSCC)remain largely undefined.This study aimed to identify critical factors and elucidate the epigenetic mechanism...Background:The regulatory mechanisms governing vasculogenic mimicry(VM)in oral squamous cell carcinoma(OSCC)remain largely undefined.This study aimed to identify critical factors and elucidate the epigenetic mechanisms underlying VM in OSCC.Methods:Bioinformatics analysis was performed utilizing single-cell RNA-seq,bulk RNA-seq,and histone H3 lysine 27 acetylation(H3K27ac)Chromatin Immunoprecipitation(ChIP)-seq data obtained from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases.ChIP-qPCR was used to validate the binding of ETS transcription factor ELK4(ELK4)to the dihydrofolate reductase(DHFR)enhancer.In vitro VM formation and invasion of OSCC cells were assessed using Matrigel-based tube formation and Transwell assays,respectively.Results:Elevated expression of VM-related genes predicts unfavorable prognosis in OSCC patients.High-dimensional weighted gene co-expression network analysis(hdWGCNA)identified epithelial subcluster C4 as most strongly associated with VM and metastasis.Three co-expression modules within this subcluster exhibited significant positive correlations with both phenotypic traits.Among the 30 eigengenes from the three modules,DHFR emerged as a key regulator of VM and metastasis.Knockdown or inhibition of DHFR significantly suppressed VM formation and invasion in OSCC cells.Mechanistically,ELK4 activated DHFR transcription through direct binding to its enhancer.DHFR overexpression rescued VM and invasion impairment induced by ELK4 knockdown.Conclusion:DHFR was a pivotal enhancer-regulated gene driving VM and metastasis in OSCC.ELK4 directly binds to DHFR enhancer regions to activate its transcription,thereby promoting these malignant phenotypes.These findings identified the ELK4/DHFR axis as a promising therapeutic target for anti-angiogenic intervention in OSCC.展开更多
Phytomelatonin,an emerging plant hormone,plays vital roles in plant growth,development,and stress adaptation(Arnao et al.,2022;Ullah et al.,2024).It acts both as a direct antioxidant and a signaling molecule,engaging ...Phytomelatonin,an emerging plant hormone,plays vital roles in plant growth,development,and stress adaptation(Arnao et al.,2022;Ullah et al.,2024).It acts both as a direct antioxidant and a signaling molecule,engaging complex networks and interacting with other phytohormones(Liu et al.,2022;Khan et al.,2023).Although phytomelatonin receptors(PMTRs)have been identified in many plants(Wei et al.,2018;Wang et al.,2022;Liu et al.,2025),the downstream signaling mechanisms,particularly receptor-mediated protein modifications and transcriptional regulation,remain poorly characterized.展开更多
Tooth morphogenesis is orchestrated by a complex interplay of signaling pathways and transcription factors that control cell proliferation,apoptosis,and differentiation,with the Wnt/β-catenin signaling pathway playin...Tooth morphogenesis is orchestrated by a complex interplay of signaling pathways and transcription factors that control cell proliferation,apoptosis,and differentiation,with the Wnt/β-catenin signaling pathway playing a pivotal role.However,the comprehensive regulatory mechanisms of Wnt/β-catenin signaling remain largely unclear.Smad7,a key antagonist of the TGF-βsuperfamily,is essential for maintaining tissue homeostasis and ensuring proper cellular function.Our previous study has demonstrated that Smad7 knockout in mice leads to impaired proliferative property of tooth germ cells,resulting in small molars.Here,we identified SMAD7 expression in human dental papilla and dental pulp,colocalized with β-CATENIN and cell proliferationrelated proteins.RNA sequencing analysis revealed a significant reduction in Wnt signaling activity in Smad7-deficient mouse tooth germs.Using lentivirus transfection,we established SMAD7-knockdown human dental papilla stem cells,which manifested remarkably blunt proliferation rate,along with diminished Wnt signaling activity.In vivo transplantation investigations further revealed the indispensable role of SMAD7 in dentin formation.Mechanistically,we revealed that β-CATENIN interacts with P-SMAD2/3 and SMAD7 through co-immunoprecipitation and yeast two-hybrid assays.Inhibition of TGF-β pathway or disruption of SMAD7/β-CATENIN transcription factor complex formation potently impacted Wnt/β-catenin activities,indicating both direct and indirect regulatory mechanisms.These findings highlight the critical role of SMAD7 in the proliferation and diffe rentiation of human dental stem cells,which could contribute to dental tissue regeneration and engineering.展开更多
ERD4 proteins,members of the early responsive-to-dehydration family,act as plasma membrane ion channels that contribute to ion homeostasis and modulate plant response to abiotic stresses.However,the functions of ERD4 ...ERD4 proteins,members of the early responsive-to-dehydration family,act as plasma membrane ion channels that contribute to ion homeostasis and modulate plant response to abiotic stresses.However,the functions of ERD4 homologs in non-vascular species remain largely unexplored.Here,we characterized an ERD4 family homolog in Physcomitrium patens(Hedw.)Mitt.,PpCSC1(Calcium-permeable Stress-responsive Cation Channel 1),and investigated its role in salt stress response.PpCSC1 localized to the plasma membrane and functioned as a non-selective cation channel permeable to Na^(+),K^(+),Ca^(2+),and Mg^(2+).Under salt treatment,PpCSC1 transcripts were markedly downregulated,whereas overexpression lines exhibited enhanced salt sensitivity.Ion content analysis further revealed reduced K^(+)accumulation,lowered K^(+)/Na^(+)ratios,and elevated Mg^(2+)levels,collectively disrupting ionic homeostasis and impairing salt tolerance.Transcriptional regulation analysis revealed that the C2H2-type zinc finger transcription factor PpSTOP2 directly activated PpCSC1 expression.Notably,PpSTOP2 knockout plants displayed reduced PpCSC1 mRNA accumulation and improved salt tolerance.Together,these findings indicate that PpCSC1 is a plasma membrane-localized cation channel that negatively regulates salt tolerance by disturbing ion balance,and that its regulation by PpSTOP2 integrates upstream signaling with downstream physiological responses.This work provides new insight into how non-selective ion channels shape stress adaptation in early land plants.展开更多
Erysiphe necator is a destructive fungal pathogen that compromises grapevine yield and quality,leading to substantial economic losses.Therefore,elucidating host resistance mechanisms is essential.In this study,we iden...Erysiphe necator is a destructive fungal pathogen that compromises grapevine yield and quality,leading to substantial economic losses.Therefore,elucidating host resistance mechanisms is essential.In this study,we identified an ethylene response factor,VqERF1B,that exhibits sustained high expression during E.necator infection in Chinese wild grape Vitis quinquangularis accession ‘Danfeng-2'.Transient overexpression of VqERF1B in grape leaves enhanced resistance to E.necator by elevating transcript levels of pathogenesis-related(PR) genes,including PR1,PR2,PR5,and PR10.Conversely,silencing VqERF1B resulted in increased susceptibility.Moreover,transgenic Arabidopsis lines stably overexpressing VqERF1B exhibited enhanced resistance to powdery mildew,associated with elevated PR gene expression and increased accumulation of reactive oxygen species(ROS).A series of assays identified VqMAPK3,a phosphorylated mitogen-activated protein kinase,as a direct interactor of VqERF1B.Furthermore,VqERF1B was shown to bind directly to the promoters of VqPRs,thereby activating their transcription.Notably,the VqMAPK3-VqERF1B complex exhibited greater transactivation activity on VqPR promoters than VqERF1B alone,indicating that VqMAPK3 positively modulates VqERF1Bmediated transcription of PR genes.This work advances understanding of the molecular basis of grape resistance to E.necator and provides a foundation for molecular breeding strategies.展开更多
Spared regions of the damaged central nervous system undergo dynamic remodelling and exhibit a remarkable potential for therapeutic exploitation1.Lesion-remote astrocytes(LRAs),which interact with viable neurons and g...Spared regions of the damaged central nervous system undergo dynamic remodelling and exhibit a remarkable potential for therapeutic exploitation1.Lesion-remote astrocytes(LRAs),which interact with viable neurons and glia,undergo reactive transformations whose molecular and functional properties are poorly understood2.Here,using multiple transcriptional profiling methods,we investigated LRAs from spared regions of mouse spinal cord following traumatic spinal cord injury.展开更多
Objective:Osteoarthritis(OA)is a degenerative joint disease characterized by extracellular matrix(ECM)degradation,chondrocyte apoptosis,and chronic inflammation.Cartilage destruction and ECM degeneration contribute to...Objective:Osteoarthritis(OA)is a degenerative joint disease characterized by extracellular matrix(ECM)degradation,chondrocyte apoptosis,and chronic inflammation.Cartilage destruction and ECM degeneration contribute to joint function loss and disability.Signal transducer and activator of transcription 3(STAT3)up-regulates the expression of MMP-13,which degrades collagen Ⅱ.Our previous study found that 5,7,3',4'-tetramethoxyflavone(TMF)exhibited protective effects on OA chondrocytes.This study aims to investigate the protective role of TMF in inhibiting ECM degradation by mediating the Sirt1/STAT3 signaling pathway.Methods:Rat OA models were established by the injection of monosodium iodoacetate(MIA).Hematoxylin&eosin(HE)staining and immunohistochemistry(IHC)analysis were performed.IL-1β stimulated C28/I2 cells were used as OA-like chondrocyte cell model.Western blotting assays were used to determine the protein expression.Results:The expression of MMP-13 was upregulated while type Ⅱ collagen expression is downregulated,and the phosphorylation level of STAT3 is increased in rat OA models.TMF reverses the STAT3-mediated expression of MMP-13 and type v collagen.Activation of STAT3 or inhibition of Sirt1 function attenuates the inhibitory effect of TMF on ECM degradation.Conclusion:TMF can inhibit ECM degradation mediated by the STAT3 signal pathway by activating Sirt1 expression in OA cell and animal models.展开更多
Objectives Dysregulated osteoclast function contributes to skeletal diseases.However,the specific ubiquitination regulators of the osteoclastogenesis repressor MafB,particularly at the post-translational level,remain ...Objectives Dysregulated osteoclast function contributes to skeletal diseases.However,the specific ubiquitination regulators of the osteoclastogenesis repressor MafB,particularly at the post-translational level,remain undefined.This study aims to identify ubiquitin-specific proteases(USPs)that deubiquitinate MafB and enhance its stability.Methods We constructed a MafB-conjugated luciferase and overexpressed 40 individual USPs,measuring changes in luciferase activity.The identified USP was overexpressed in human CD14^(+) peripheral blood mononuclear cells(PBMCs)to evaluate its effect.Osteoclast differentiation was assessed through osteoclast marker Integrin alpha-V(CD51)staining and Western blot analysis.Co-immunoprecipitation(co-IP)was performed to assess the interplay.The influence on MafB ubiquitination and degradation was evaluated via immunoprecipitation and Western blot.Finally,MafB was knocked down in the USP-overexpressing PBMCs to analyze its effect on osteoclast differentiation.Results Overexpression of ubiquitin-specific protease 29(USP29)significantly increased MafB expression by approximately 75%(p<0.0001).Elevated USP29 levels strongly inhibited osteoclastic differentiation in CD14^(+) PBMCs(p<0.0001).USP29 was found to interact with MafB,markedly reducing its ubiquitination and subsequent degradation in PBMCs(p<0.001).Knocking down MafB in USP29-overexpressing PBMCs alleviated the inhibitory effect of USP29 on osteoclastogenesis.Conclusion USP29 acts as a potent stabilizer of MafB,inhibiting osteoclastogenesis in human CD14^(+) PBMCs,at least in part,by enhancing MafB stability.These findings expand our understanding of USP29’s role and the post-translational regulation of MafB.Furthermore,USP29 serves as a vital factor that controls osteoclast differentiation,and its regulatory function is at least partially mediated by deubiquitinating and stabilizing MafB.展开更多
基金supported by the National Natural Science Foundation of China(31872706)the National Key Research and Development Program of China(2019 YFD1000603).
文摘Zanthoxylum bungeanum is an economically important crop worldwide due to its high content of aroma-producing monoterpenoids,and development of varieties with enhanced flavor and overall quality is a crucial research area.However,the transcriptional regulatory mechanisms underlying monoterpenoid synthesis in Z.bungeanum remain unclear,hindering these breeding efforts.In this study,RNA sequencing,gas chromatography–mass spectrometry,and other molecular biology techniques were used to identify the underlying transcriptional regulation mechanisms.Two transcription factors,ZbbHLH2 and ZbERF6,were identified as key regulators of monoterpenoid synthesis in Z.bungeanum that upregulate various monoterpenoid synthesis-associated genes and are novel transcriptional activators of ZbIDI,which encodes the rate-limiting enzyme in plant monoterpenoid synthesis.Functional analysis revealed that the expression of three genes[1]modulates monoterpenoid accumulation in Z.bungeanum peel.These findings provide novel insights into the metabolic regulatory network of monoterpenoid synthesis in Z.bungeanum peel,offer potential strategies for the biofortification of specific monoterpenoids,and will promote the development of Z.bungeanum germplasm for targeted breeding and quality improvement.
基金supported by the National Natural Science Foundation of China(32372641)the Shandong Provincial Natural Science Foundation,China(ZR2024QC143)the Liaoning Provincial Natural Science Foundation,China(2022-MS-258)。
文摘Anthocyanins are important flavonoid pigments in the coloration of fruits.To identify candidate genes involved in anthocyanin accumulation,metabolic and transcriptome analyses were conducted in‘Nanguo'pear and its red sport cultivar‘Nanhong'pear.The results showed that‘Nanhong'pear had significantly higher anthocyanin and flavonol contents.Additionally,transcriptomic analysis showed that there were significant differences in the expression of genes involved in phenylpropanoid and flavonoid biosynthesis pathways between the two cultivars,with PuGSTF12 being the most upregulated gene in the‘Nanhong'cultivar.Further analysis identified a novel MYB transcription factor(PuMYB93),and its silencing repressed PuGSTF12 expression and anthocyanin accumulation,suggesting it plays an essential role in the regulation of anthocyanin biosynthesis.Moreover,yeast one-hybrid analysis,electrophoretic mobility shift assay,andβ-glucuronidase assay indicated that PuMYB93 can directly bind to the PuGSTF12 promoter to positively regulate its expression.Additionally,PuGSTF12 silencing suppressed the coloration of PuMYB93-OE peels,suggesting that PuGSTF12 act downstream of PuMYB93.Overall,the findings of this study show that PuMYB93 promotes anthocyanin transport in pears by regulating PuGSTF12 expression to further enhance anthocyanin accumulation.
基金supported by the SECIHTI project Ciencia Basica y de Frontera(No.CBF2023-2024-1141)https://secihti.mx/(accessed on 01 August 2025).
文摘Objectives:Oxidative stress(OS)plays a pivotal role in chronic and neurodegenerative diseases,which has sparked interest in molecules that modulate redox-regulating enzymes.Melatonin and its metabolites exhibit antioxidant properties;however,their molecular mechanisms of enzymatic and transcriptional modulation remain unclear.This study aimed to investigate,through an exploratory in silico approach,the interactions of melatonin and related compounds with OS-related enzymes to generate hypotheses about their role in cellular redox control.Methods:A rational selection of antioxidant,pro-oxidant,and transcriptional targets was performed.Ligands were optimized at the DFT level(M05-2X/6-311+G(d,p))and docked to OS related enzymes.Docking results were analyzed using polygenic antioxidant indices(PAOX)and a similarity interaction index(SSI).Molecular dynamics simulations of selected complexes provided additional insight into potential ligand-protein interaction mechanisms.Results:In silico analyses revealed that N1-acetyl-5-methoxykynuramine(AMK),N1-acetyl-N2-formyl-5-methoxykynuramine(AFMK),and 3-hydroxymelatonin(3OH-M)could partially inhibit pro-oxidant enzymes such as neuronal nitric oxide synthase(nNOS),5-lipoxygenase(5-LOX),thioredoxin reductase(TrxR),and nicotinamide adenine dinucleotide phosphate oxidase(NOX5).The N-(2-(2-acetyl-6,7-dihydroxy-1H-indol-3-yl)ethyl)acetamide(IIcD)and N-(2-(6-hydroxy-7-mercapto-5-methoxy-1H-indol)ethyl)acetamide(dM38)derivatives could potentially stabilize superoxide dismutase(SOD1)and catalase(CAT)enzymes,respectively.Finally,AFMK and dM38 showed consistent interactions with transcriptional regulators,particularly peroxisome proliferator-activated receptor alpha(PPARα)and Kelchlike ECH-associated protein 1(KEAP1).Conclusion:These studies about melatonin-related compounds support a multifactorial profile of redox modulation and provide mechanistic hypotheses for future experimental validation.Among these approaches,the interaction-similarity index is introduced as a novel tool to facilitate the identification of promising redox-active candidates.
基金supported by the China Agricultural Research System(Grant No.CARS-09)the Central Government Guiding Local Science and Technology Development Project(Grant No.YDZX2023029)the Gansu Planning Projects on Science and Technology(Grant No.23CXNJ0013).
文摘Flavonoids,abundant in the fruits,are pivotal to their growth,development,and storage.In addition,they have significant beneficial effects on human health.Consequently,research is increasingly concentrating on the regulatory mechanisms governing flavonoid biosynthesis in fruits.Phytohormones are involved in the regulation of flavonoid biosynthesis.The abscisic acid,ethylene,jasmonic acid,cytokinins,and brassinosteroids promote flavonoid biosynthesis,while auxin negatively regulates flavonoid biosynthesis.Subsequently,transcription factors from the MYB,bHLH,WRKY,NAC,and bZIP families are pivotal in regulating flavonoid biosynthesis.In addition,non-coding RNAs(microRNA and lncRNA)also participate in the regulation of flavonoids biosynthesis.MicroRNAs are generally believed to negatively regulate flavonoid metabolism in fruits,while lncRNAs have the opposite effect.Furthermore,the interactions between plant hormones,transcription factors,and non-coding RNAs in fruit flavonoid biosynthesis were analyzed.Ultimately,a foundational regulatory network for fruit flavonoid biosynthesis was hereby established.
基金supported by the National Key R&D Program of China(Grant No.2022YFD1600700)Natural Science Foundation of China(Grant Nos.32372644,32472661)+1 种基金Natural Science Foundation of Universities in Anhui Province,China(Grant Nos.2022AH050931,2023AH051043)Anhui Provincial Natural Science Foundation,China(Grant No.2108085MC105).
文摘Anthocyanins are the main pigments in ripe strawberry fruits.FaMYB10 and abscisic acid(ABA)are the two major regulators of anthocyanin biosynthesis in the maturation process of strawberry fruits.However,the transcriptional regulation of FaMYB10,ABA biosynthesis,and anthocyanin accumulation in strawberry(Fragaria×ananassa)remain largely unknown.Here,a yeast one-hybrid screen using the FaMYB10 promoter identified a class B MADS-domain transcription factor,FaMADS6 in strawberry.FaMADS6 exhibited high expression at the early developmental stage but was hardly detected during maturation of strawberry fruit,a pattern opposite to accumulation of anthocyanins.Transcriptional upregulation of FaMADS6 repressed anthocyanin accumulation and expression of FaMYB10 and the anthocyanin biosynthetic genes,FaCHS,FaCHI,FaF3H,FaANS,and FaUFGT.In contrast,downregulation of FaMADS6 promoted the expression of FaMYB10 and the anthocyanin biosynthetic genes.The promoters of the anthocyanin biosynthetic genes were not directly bound by FaMADS6,in contrast to FaMYB10.Analysis of the DNA binding sequences of FaMADS6 revealed that it also interacted with the promoters of FaNCED2 and FaPYR1,which are involved in the biosynthesis and perception of ABA.Overexpression of FaMADS6 significantly suppressed FaNCED2 and FaPYR1 and ABA synthesis in transgenic strawberry.Together,our findings suggest that FaMADS6 functions as a suppressor of anthocyanin accumulation by directly downregulating FaMYB10 and ABA production during strawberry fruit maturation.
基金supported by the Guizhou Provincial Basic Research Program(Natural Science)[(2024)648]the Program of China National Tobacco Corporation(110202101032(JY-09),110202201003(JY-03))+2 种基金the Program of Guizhou Branch of China National Tobacco Corporation(2023XM02,2022XM05 and 2024XM01)the Qiankehe Platform Project(ZSYS[2025]028)the Program of China National Tobacco Corporation(110202102034).
文摘Tobacco(Nicotiana tabacum,2n=48)is a key non-food economic crop,yet its stress response and gene regulatory mechanisms remain poorly understood.By analyzing 603 transcriptome datasets,this study identified 1405 tissue-specific genes,revealing tissue-specific synthesis of terpenoids and other ecologically important secondary metabolites in sepals and other tissues.Comparative stress-response analysis highlighted distinct gene expression patterns in leaves and roots under biotic and abiotic stresses.Additionally,28,396 expression quantitative trait loci(eQTLs)were mapped in leaves,offering valuable genetic regulatory markers.These findings provide crucial insights into tobacco’s gene expression characteristics and their functional implications,serving as a foundation for future research.
基金the Projects of Science and Technology of Shanghai(22N11900400)Key Projects of Beijing Institute of Life Science and Technology(2024400CB0050)+1 种基金National Natural Science Foundation of China(32488102)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA24030303)。
文摘Flavonoids are crucial secondary metabolites widely distributed in plants,playing vital roles in diverse biological processes.Although the flavonoid biosynthesis pathway has been extensively characterized,the transcriptional regulatory mechanisms remain poorly understood.In this study,we identify the miR166–ATHB14-LIKE module comprising the miR166 and its target gene ATHB14-LIKE as a key regulator of flavonoid biosynthesis in soybean(Glycine max).Knockdown of miR166 or overexpression of ATHB14-LIKE upregulated multiple flavonoid biosynthesis genes,leading to increased flavonoid accumulation.Conversely,miR166 overexpression suppressed these genes and reduced flavonoid levels.We further show that ATHB14-LIKE directly activates specific flavonoid biosynthesis genes by binding to their promoters.Additionally,ATHB14-LIKE forms homodimers and heterodimers with homologous proteins to regulate downstream flavonoid biosynthesis genes.Together,our findings demonstrate that the miR166–ATHB14-LIKE module controls soybean flavonoid content by coordinating the expression of key biosynthetic genes.
基金supported by the National Natural Science Foundation of China(Grant Nos.32102311 and 32102338)the China Postdoctoral Science Foundation(Grant No.2021M690129).
文摘Drought stress significantly impedes apple growth,development,and yield,leading to substantial economic losses within the global apple industry.Malus prunifolia(Mp),a commonly utilized apple rootstock,has shown promise in augmenting cultivated apple resistance to abiotic stress.Although Alfin-like(ALs)proteins have demonstrated pivotal roles in dicotyledonous plants'response to abiotic stresses,knowledge about AL genes in apple rootstocks is limited,and their functions remain largely elusive.In this study,we identified and characterized 10 MpAL gene members in the apple rootstock genome,confirming their localization within the nucleus.Our investigation revealed the significant regulation of MpALs'expression under drought and abscisic acid(ABA)stresses in M.prunifolia.In this study,one of the members,MpAL1,was selected for further exploration in Arabidopsis and apple to explore its potential function in response to drought and ABA stresses.The results showed that overexpression-MpAL1 transgenic apple calli grew significantly better than WT and MpAL1-RNAi lines,which regulates the accumulation of H_(2)O_(2)and O_(2).-levels.Additionally,transgenic Arabidopsis plants overexpressing MpAL1 exhibited positively regulating antioxidant enzymes activities under stress treatments.Further study showed that silencing MpAL1 in apple plants showed obvious chlorosis in leaves,and accumulation of reactive oxygen species under drought stress.Moreover,our detailed analysis established that MpAL1 regulates several drought and ABA-responsive genes,exerting an influence on their expression in transgenic apple.Collectively,our findings identify MpAL1 as a positive regulator that increases drought stress in apple,shedding light on its potential significance in bolstering drought resistance in this fruit crop.
基金funding from the Natural Science Foundation of Sichuan Province,China(Grant Nos.2025ZNSFSC1100,2024NSFSC0324)the National Key Research and Development Project(Grant No.2017YFC0505104)+2 种基金the Sichuan Agricultural University Dual Support Plan Special Project(Grant No.2024ZYTS020)the Postdoctoral Fellowship Program(Grade B)of China Postdoctoral Science Foundation(Grant No.GZB20250863)the China Postdoctoral Science Foundation(Grant No.2025M773740).
文摘Anthocyanins play a crucial role in shaping the visual appeal and nutritional quality of fruits.Previous research on anthocyanin biosynthesis in sweet cherry(Prunus avium L.)has primarily relied on single-omics approaches or focused on a limited range of metabolites,leaving the regulatory mechanisms and dynamic metabolism of anthocyanins during ripening inadequately characterized.This study integrated anthocyanin-targeted metabolomics and transcriptomics to identify key anthocyanins in sweet cherry and construct a transcriptional regulatory network for anthocyanin biosynthesis.A novel bHLH transcription factor,Prunus avium bHLH transcription factor 102(PavbHLH102),was identified,and its role in regulating cyanidin levels was validated through overexpression and silencing experiments.Both in vitro and in vivo assays demonstrated that PavbHLH102 activates key anthocyanin biosynthetic genes,including PavF3H,PavDFR,and PavUFGT,thereby enhancing fruit coloration.Notably,PavF3′H upregulation significantly increased cyanidin accumulation.This study provides new insights into anthocyanin regulation in sweet cherry and offers valuable resources for improving fruit quality.
基金funded by the National Natural Science Foundation of China(Grant Nos.82204517 to T.Z.and 82404756 to J.Z.)the Science and Technology Program in Medicine and Health of Zhejiang Province(Grant No.2023KY726 to T.Z.).
文摘Background:Aberrant expression of transcription factors(TFs)is a key mechanism mediating tumor immune escape and therapeutic resistance.The involvement of E26 transformation-specific(ETS)family of TFs in immune regulation is not fully understood.The study aimed to elucidate the function of E-twenty-six variant 4(ETV4)in tumor immune evasion and its potential as a predictive biomarker for immunotherapy in melanoma.Methods:The expression patterns of ETS family TFs were analyzed in melanoma and hepatocellular carcinoma(HCC).Single-cell RNA sequencing(scRNA-seq)was used to dissect the cellular expression and function of ETV4 in the tumor microenvironment.Functional studies and murine models were employed to investigate the role of ETV4 in T cell-mediated tumor killing and tumor growth.The correlation between ETV4 expression level and patient responsiveness to programmed cell death protein 1(PD-1)blockade therapy was evaluated.Results:TFs in the ETS family were found to effectively stratify melanoma and HCC patients into prognostic subgroups.In melanoma,the polyoma enhancer activator 3(PEA3)subfamily,particularly ETV4 and ETV5,showed a negative correlation with immune infiltration.scRNA-seq analysis showed that ETV4 is preferentially expressed in melanoma cells and involves in mediating tumor-immunocyte interactions.Functional studies demonstrated that ETV4 impairs T cell-mediated tumor killing by transcriptionally upregulating programmed death-ligand 1(PD-L1).In immunocompetent murine models,ETV4 downregulation significantly suppressed tumor growth.Furthermore,high ETV4 expression correlated with poor responses to anti-PD-1 therapy.Conclusion:Our findings identify ETV4 as a key transcriptional regulator of immune evasion in melanoma by controlling PD-L1 expression.ETV4 may act as a predictive biomarker for immunotherapy outcomes.
基金supported by the National Natural Science Foundation of China(Grant Nos.31870695,32071828)the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Flower color is an essential trait in ornamental plant breeding. Lycoris longituba is a popular ornamental plant native to central eastern China. The decrease in anthocyanin accumulation causes L. longituba petal color fading during flower development, which considerably affects the ornamental value of L. longituba. However, mechanisms underlying anthocyanin biosynthesis inhibition during L. longituba petal development remain unclear. In this study, three LlDFR genes were confirmed to be involved in anthocyanin biosynthesis and LlDFRc exerted the strongest promoting effect on anthocyanin accumulation. According to the correlation analysis results, LlbHLH12 exhibited the strongest negative correlation with LlDFRc. Quantitative real-time PCR analysis showed that LlbHLH12 was highly expressed during the medium bud and full bloom stages of flower development. LlbHLH12 was identified as a member of subgroup XII of bHLH transcription factor family. Subcellular localization and transcriptional activation ability assay revealed that LlbHLH12 was located in the nucleus without transcriptional activation activity. Overexpression of LlbHLH12 in Nicotiana tabacum and L. longituba inhibited anthocyanin accumulation by suppressing the expression of anthocyanin biosynthetic pathway genes. Furthermore, yeast one-hybrid, dual-luciferase, and β-glucuronidase activity assays showed that LlbHLH12 directly bound to the promoters of LlPAL and LlDFRc and suppressed their expression to inhibit anthocyanin biosynthesis. Overall, our study identified a novel bHLH repressor negatively regulating anthocyanin biosynthesis and provided new insights into the molecular mechanisms underlying color fading in L. longituba petals.
基金supported by grants from Shanghai Agriculture Applied Technology Development Program(Grant No.2022-02-08-00-12-F01109)the National Natural Science Foundation of China(Grant No.32272721).
文摘The B-box(BBX)gene family plays a vital role in plant growth,development,and stress responses.This study aimed to characterize the SmBBX gene family in eggplant(Solanum melongena L.),addressing the lack of systematic bioinformatics and functional studies in this species.A total of 33 SmBBX genes were identified through genome-wide analysis.These genes were phylogenetically grouped into five major clades,with shared domain structures,motifs,and genomic architectures among clade members.The gene duplication analysis revealed segmental duplication as the primary mechanism underlying the expansion of SmBBX proteins in eggplant.Additionally,expression profiling across diverse tissues and abiotic stress conditions,combined with the construction of protein—protein interaction networks and luciferase complementation assay,provided valuable insights into the functional roles of SmBBX genes.SmBBX21-2 and SmBBX22 were identified as the key regulators of anthocyanin biosynthesis,activating the expression of SmCHS and SmDFR promoters.Functional validation via heterologous and homologous overexpression demonstrated that SmBBX22 promoted anthocyanin accumulation by upregulating the expression of structural genes(SmCHS,SmF3H,SmF3′5′H,SmDFR,and SmANS)and transcription factors(SmTT8 and SmHY5)important for anthocyanin biosynthesis.Furthermore,the integration of DNA affinity purification sequencing and RNA-seq data revealed the direct transcriptional targets of SmBBX22,including genes involved in secondary metabolism,hormone signaling,and developmental regulation.This highlighted the role of SmBBX22 in phenylpropanoid and flavonoid biosynthesis.This study lays the foundation for understanding the functional roles of BBX genes in eggplant and provides new directions for future research in plant metabolism and stress adaptation.
基金supported by Hebei Natural Science Foundation(H2024206476)Medical Science Research Project of Hebei(20240101).
文摘Background:The regulatory mechanisms governing vasculogenic mimicry(VM)in oral squamous cell carcinoma(OSCC)remain largely undefined.This study aimed to identify critical factors and elucidate the epigenetic mechanisms underlying VM in OSCC.Methods:Bioinformatics analysis was performed utilizing single-cell RNA-seq,bulk RNA-seq,and histone H3 lysine 27 acetylation(H3K27ac)Chromatin Immunoprecipitation(ChIP)-seq data obtained from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases.ChIP-qPCR was used to validate the binding of ETS transcription factor ELK4(ELK4)to the dihydrofolate reductase(DHFR)enhancer.In vitro VM formation and invasion of OSCC cells were assessed using Matrigel-based tube formation and Transwell assays,respectively.Results:Elevated expression of VM-related genes predicts unfavorable prognosis in OSCC patients.High-dimensional weighted gene co-expression network analysis(hdWGCNA)identified epithelial subcluster C4 as most strongly associated with VM and metastasis.Three co-expression modules within this subcluster exhibited significant positive correlations with both phenotypic traits.Among the 30 eigengenes from the three modules,DHFR emerged as a key regulator of VM and metastasis.Knockdown or inhibition of DHFR significantly suppressed VM formation and invasion in OSCC cells.Mechanistically,ELK4 activated DHFR transcription through direct binding to its enhancer.DHFR overexpression rescued VM and invasion impairment induced by ELK4 knockdown.Conclusion:DHFR was a pivotal enhancer-regulated gene driving VM and metastasis in OSCC.ELK4 directly binds to DHFR enhancer regions to activate its transcription,thereby promoting these malignant phenotypes.These findings identified the ELK4/DHFR axis as a promising therapeutic target for anti-angiogenic intervention in OSCC.
基金supported by the grants from the Key Research and Development Program of Xinjiang Uygur autonomous region in China(Grant No.2023B02017)the National Key Research and Development Program of China(Grant No.2024YFD2300703)+1 种基金the financial support from the Beijing Rural Revitalization Agricultural Science and Technology Project(Grant No.NY2401080000),BAIC01-2025the 2115 Talent Development Program of China Agricultural University.
文摘Phytomelatonin,an emerging plant hormone,plays vital roles in plant growth,development,and stress adaptation(Arnao et al.,2022;Ullah et al.,2024).It acts both as a direct antioxidant and a signaling molecule,engaging complex networks and interacting with other phytohormones(Liu et al.,2022;Khan et al.,2023).Although phytomelatonin receptors(PMTRs)have been identified in many plants(Wei et al.,2018;Wang et al.,2022;Liu et al.,2025),the downstream signaling mechanisms,particularly receptor-mediated protein modifications and transcriptional regulation,remain poorly characterized.
基金supported by the National Key Research and Development Program of China to W.Tian (2022YFA1104400)the National Natural Science Foundation of China to T.Chen (82100959)a grant from the Sichuan Science and Technology Program to Z.Liu (2024YFFK0068)。
文摘Tooth morphogenesis is orchestrated by a complex interplay of signaling pathways and transcription factors that control cell proliferation,apoptosis,and differentiation,with the Wnt/β-catenin signaling pathway playing a pivotal role.However,the comprehensive regulatory mechanisms of Wnt/β-catenin signaling remain largely unclear.Smad7,a key antagonist of the TGF-βsuperfamily,is essential for maintaining tissue homeostasis and ensuring proper cellular function.Our previous study has demonstrated that Smad7 knockout in mice leads to impaired proliferative property of tooth germ cells,resulting in small molars.Here,we identified SMAD7 expression in human dental papilla and dental pulp,colocalized with β-CATENIN and cell proliferationrelated proteins.RNA sequencing analysis revealed a significant reduction in Wnt signaling activity in Smad7-deficient mouse tooth germs.Using lentivirus transfection,we established SMAD7-knockdown human dental papilla stem cells,which manifested remarkably blunt proliferation rate,along with diminished Wnt signaling activity.In vivo transplantation investigations further revealed the indispensable role of SMAD7 in dentin formation.Mechanistically,we revealed that β-CATENIN interacts with P-SMAD2/3 and SMAD7 through co-immunoprecipitation and yeast two-hybrid assays.Inhibition of TGF-β pathway or disruption of SMAD7/β-CATENIN transcription factor complex formation potently impacted Wnt/β-catenin activities,indicating both direct and indirect regulatory mechanisms.These findings highlight the critical role of SMAD7 in the proliferation and diffe rentiation of human dental stem cells,which could contribute to dental tissue regeneration and engineering.
基金supported by the National Natural Science Foundation of China(Grant No.31970658 and No.32400208)the Zhejiang Provincial Natural Science Foundation of China(Grant No.LD24C130002 and No.LQN25C020001)the Scientific Research Foundation of China Jiliang University.
文摘ERD4 proteins,members of the early responsive-to-dehydration family,act as plasma membrane ion channels that contribute to ion homeostasis and modulate plant response to abiotic stresses.However,the functions of ERD4 homologs in non-vascular species remain largely unexplored.Here,we characterized an ERD4 family homolog in Physcomitrium patens(Hedw.)Mitt.,PpCSC1(Calcium-permeable Stress-responsive Cation Channel 1),and investigated its role in salt stress response.PpCSC1 localized to the plasma membrane and functioned as a non-selective cation channel permeable to Na^(+),K^(+),Ca^(2+),and Mg^(2+).Under salt treatment,PpCSC1 transcripts were markedly downregulated,whereas overexpression lines exhibited enhanced salt sensitivity.Ion content analysis further revealed reduced K^(+)accumulation,lowered K^(+)/Na^(+)ratios,and elevated Mg^(2+)levels,collectively disrupting ionic homeostasis and impairing salt tolerance.Transcriptional regulation analysis revealed that the C2H2-type zinc finger transcription factor PpSTOP2 directly activated PpCSC1 expression.Notably,PpSTOP2 knockout plants displayed reduced PpCSC1 mRNA accumulation and improved salt tolerance.Together,these findings indicate that PpCSC1 is a plasma membrane-localized cation channel that negatively regulates salt tolerance by disturbing ion balance,and that its regulation by PpSTOP2 integrates upstream signaling with downstream physiological responses.This work provides new insight into how non-selective ion channels shape stress adaptation in early land plants.
基金This study is granted by the National Natural Science Foundation of China(52209055,52379041,and 32272667)the Yunnan Fundamental Research Projects,China (202401AU070197,202501AW070013,202501BC070015,and 202501AT070377)+3 种基金the Yunnan Education Department Project,China (2024J0079)the Kunming University of Science and Technology Talent Development Project,China (KKZ3202423161)the Yunnan Key Laboratory of Efficient Utilization of Agricultural Water Resources and Intelligent Control,China (202449CE340014)the Yunnan Intelligent Water-Fertilizer-Pesticide Integration Technology and Equipment Innovation Team,China (202505AS350025)。
文摘Erysiphe necator is a destructive fungal pathogen that compromises grapevine yield and quality,leading to substantial economic losses.Therefore,elucidating host resistance mechanisms is essential.In this study,we identified an ethylene response factor,VqERF1B,that exhibits sustained high expression during E.necator infection in Chinese wild grape Vitis quinquangularis accession ‘Danfeng-2'.Transient overexpression of VqERF1B in grape leaves enhanced resistance to E.necator by elevating transcript levels of pathogenesis-related(PR) genes,including PR1,PR2,PR5,and PR10.Conversely,silencing VqERF1B resulted in increased susceptibility.Moreover,transgenic Arabidopsis lines stably overexpressing VqERF1B exhibited enhanced resistance to powdery mildew,associated with elevated PR gene expression and increased accumulation of reactive oxygen species(ROS).A series of assays identified VqMAPK3,a phosphorylated mitogen-activated protein kinase,as a direct interactor of VqERF1B.Furthermore,VqERF1B was shown to bind directly to the promoters of VqPRs,thereby activating their transcription.Notably,the VqMAPK3-VqERF1B complex exhibited greater transactivation activity on VqPR promoters than VqERF1B alone,indicating that VqMAPK3 positively modulates VqERF1Bmediated transcription of PR genes.This work advances understanding of the molecular basis of grape resistance to E.necator and provides a foundation for molecular breeding strategies.
文摘Spared regions of the damaged central nervous system undergo dynamic remodelling and exhibit a remarkable potential for therapeutic exploitation1.Lesion-remote astrocytes(LRAs),which interact with viable neurons and glia,undergo reactive transformations whose molecular and functional properties are poorly understood2.Here,using multiple transcriptional profiling methods,we investigated LRAs from spared regions of mouse spinal cord following traumatic spinal cord injury.
基金Project Supported by Jiangxi Provincial Natural Science Foundation(20212ACB206002)。
文摘Objective:Osteoarthritis(OA)is a degenerative joint disease characterized by extracellular matrix(ECM)degradation,chondrocyte apoptosis,and chronic inflammation.Cartilage destruction and ECM degeneration contribute to joint function loss and disability.Signal transducer and activator of transcription 3(STAT3)up-regulates the expression of MMP-13,which degrades collagen Ⅱ.Our previous study found that 5,7,3',4'-tetramethoxyflavone(TMF)exhibited protective effects on OA chondrocytes.This study aims to investigate the protective role of TMF in inhibiting ECM degradation by mediating the Sirt1/STAT3 signaling pathway.Methods:Rat OA models were established by the injection of monosodium iodoacetate(MIA).Hematoxylin&eosin(HE)staining and immunohistochemistry(IHC)analysis were performed.IL-1β stimulated C28/I2 cells were used as OA-like chondrocyte cell model.Western blotting assays were used to determine the protein expression.Results:The expression of MMP-13 was upregulated while type Ⅱ collagen expression is downregulated,and the phosphorylation level of STAT3 is increased in rat OA models.TMF reverses the STAT3-mediated expression of MMP-13 and type v collagen.Activation of STAT3 or inhibition of Sirt1 function attenuates the inhibitory effect of TMF on ECM degradation.Conclusion:TMF can inhibit ECM degradation mediated by the STAT3 signal pathway by activating Sirt1 expression in OA cell and animal models.
文摘Objectives Dysregulated osteoclast function contributes to skeletal diseases.However,the specific ubiquitination regulators of the osteoclastogenesis repressor MafB,particularly at the post-translational level,remain undefined.This study aims to identify ubiquitin-specific proteases(USPs)that deubiquitinate MafB and enhance its stability.Methods We constructed a MafB-conjugated luciferase and overexpressed 40 individual USPs,measuring changes in luciferase activity.The identified USP was overexpressed in human CD14^(+) peripheral blood mononuclear cells(PBMCs)to evaluate its effect.Osteoclast differentiation was assessed through osteoclast marker Integrin alpha-V(CD51)staining and Western blot analysis.Co-immunoprecipitation(co-IP)was performed to assess the interplay.The influence on MafB ubiquitination and degradation was evaluated via immunoprecipitation and Western blot.Finally,MafB was knocked down in the USP-overexpressing PBMCs to analyze its effect on osteoclast differentiation.Results Overexpression of ubiquitin-specific protease 29(USP29)significantly increased MafB expression by approximately 75%(p<0.0001).Elevated USP29 levels strongly inhibited osteoclastic differentiation in CD14^(+) PBMCs(p<0.0001).USP29 was found to interact with MafB,markedly reducing its ubiquitination and subsequent degradation in PBMCs(p<0.001).Knocking down MafB in USP29-overexpressing PBMCs alleviated the inhibitory effect of USP29 on osteoclastogenesis.Conclusion USP29 acts as a potent stabilizer of MafB,inhibiting osteoclastogenesis in human CD14^(+) PBMCs,at least in part,by enhancing MafB stability.These findings expand our understanding of USP29’s role and the post-translational regulation of MafB.Furthermore,USP29 serves as a vital factor that controls osteoclast differentiation,and its regulatory function is at least partially mediated by deubiquitinating and stabilizing MafB.
