Human cytomegalovirus(HCMV)is a double-strand DNA virus widely infected in human.Circular RNAs(circ RNAs)are non-coding RNAs with most functions of which keep unknown,and the effects of HCMV productive infection on ho...Human cytomegalovirus(HCMV)is a double-strand DNA virus widely infected in human.Circular RNAs(circ RNAs)are non-coding RNAs with most functions of which keep unknown,and the effects of HCMV productive infection on host circ RNA transcriptions remain unclear.In this study,we profiled 283 host circ RNAs that significantly altered by HCMV productive infection in human embryonic lung fibroblasts(HELF)by RNA deep sequencing and bioinformatics analysis.Among these,circ SP100,circ MAP3 K1,circ PLEKHM1,and circ TRIO were validated for their transcriptions and sequences.Furthermore,characteristics of circ SP100 were investigated by RT-q PCR and northern blot.It was implied that circ SP100 was produced from the sense strand of the SP100 gene containing six exons.Kinetics of circ SP100 and SP100 m RNA were significantly different after infection:circ SP100 levels increased gradually along with infection,whereas SP100 m RNA levels increased in the beginning and dropped at 24 h post-infection(hpi).Meanwhile,a total number of 257 proteins,including 10 HCMV encoding proteins,were identified potentially binding to cytoplasmic circ SP100 by RNA antisense purification(RAP)and mass spectrometry.Enrichment analysis showed these proteins were mainly involved in the spliceosome,protein processing,ribosome,and phagosome pathways,suggesting multiple functions of circ SP100 during HCMV infection.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
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
Drought stress orchestrates a phosphorylation-dependent signaling cascade that reprograms transcriptional networks to enhance crop resilience.Through a large-scale transgenic screening,we identified ZmCRK5A,a Ca^(2+)-...Drought stress orchestrates a phosphorylation-dependent signaling cascade that reprograms transcriptional networks to enhance crop resilience.Through a large-scale transgenic screening,we identified ZmCRK5A,a Ca^(2+)-independent calcium-dependent protein kinase(CDPK)-related kinase,as a master regulator of drought tolerance in maize.Mechanistically,ZmCRK5A directly phosphorylates the MYB transcriptional repressor ZmSMH4(Single MYB Histone 4)at three conserved serine residues(Ser42/43/59)within its SANT domain,as demonstrated by in vitro kinase assays and site-directed mutagenesis.This post-translational modification abolishes Zm SMH4's DNA-binding capacity to ACC cis-elements,thereby de-repressing the potassium influx channel gene Zm KCH1(K^(+)Channel 1).Functional validation revealed that Zm KCH1 overexpression confers drought resilience through optimized stomatal dynamics and water retention,whereas clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-generated zmkch1 mutants display hypersensitivity to water deficit.Crucially,field evaluations demonstrated preserved grain yield alongside enhanced drought tolerance in plants with activated Zm CRK5A-Zm SMH4-Zm KCH1 signaling.Our findings delineate a kinase-transcription factor-ion channel axis that dynamically fine-tunes drought responses while maintaining productivity,providing a strategic framework for engineering stress-adapted crops without yield penalties.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
Influenza viruses are a major cause of respiratory illness,with significant public health impact due to their ability to cause pandemics.This dialogue brought together experts including Professors George Fu Gao,Stephe...Influenza viruses are a major cause of respiratory illness,with significant public health impact due to their ability to cause pandemics.This dialogue brought together experts including Professors George Fu Gao,Stephen Cusack,Mark von Itzstein,Ervin Fodor,Jonathan Grimes,Aartjan J.W.te Velthuis,and Tao Deng to decode the pressing scientific challenges and future directions in influenza research.They discussed how structural studies of the influenza polymerase have advanced our understanding of viral RNA transcription and replication.These insights are crucial for developing new antiviral drugs,with a particular focus on targeting the polymerase and its interactions with host factors like acidic nuclear phosphoprotein 32(ANP32).The dialogue also highlighted the potential of artificial intelligence(AI)to assist in designing small-molecule drugs,offering new strategies for combating influenza.Future research will continue to unravel the complexities of the polymerase’s role in replication,aiming to translate these findings into effective therapies and resilient public health strategies.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
AIM:To investigate whether vaccinia-related kinase 1(VRK1)mediates transforming growth factor-beta2(TGF-β2)-caused epithelial-mesenchymal transition(EMT)and inflammatory responses in retinal pigment epithelial(RPE)ce...AIM:To investigate whether vaccinia-related kinase 1(VRK1)mediates transforming growth factor-beta2(TGF-β2)-caused epithelial-mesenchymal transition(EMT)and inflammatory responses in retinal pigment epithelial(RPE)cells through regulating snail family transcriptional repressor 1(SNAI1),and to validate its role in a proliferative vitreoretinopathy(PVR)mouse model.METHODS:Human RPE cell line ARPE-19 cells were treated with TGF-β2 to construct an EMT model.Western blot detected VRK1 level.The effects of VRK1 on SNAI1 expression and biological behavior of ARPE-19 cells were detected by immunofluorescence,ELISA,Transwell,and scratch assay,and the interaction between VRK1 and SNAI1 was confirmed through immunoprecipitation.A PVR mouse model was constructed,and the effects of VRK1 or/and SNAI1 on retinal damage were assessed by pathologic staining.Inflammatory factors and EMT-related proteins were assessed with ELISA and Western blot.RESULTS:VRK1 was upregulated in ARPE-19 cells after TGF-β2 treatment.Overexpression of VRK1 increased cell viability,promoted cell migration and EMT,and the levels of inflammatory factors.Silencing of VRK1 reversed the above indexes.There was a direct interaction between VRK1 and SNAI1,and overexpresssion SNAI1 weakened the impacts of silencing of VRK1.In PVR mice,silencing of VRK1 ameliorated retinal structural damage,decreased proinflammatory factor levels,and suppressed SNAI1 and mesenchymal marker expression.SNAI1 overexpression antagonized the protective effects of silencing VRK1 and exacerbated EMT and inflammatory responses.CONCLUSION:VRK1 plays a key role in retinal structural and inflammatory damage in PVR mice by regulating SNAI1 and mediating TGF-β2-caused EMT and inflammatory responses in RPE cells.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
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
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
Objective The basic helix-loop-helix(bHLH)transcription factors(TFs)are pivotal in regulating fungal growth,development,and secondary metabolism.However,the knowledge about the Ganoderma lucidum bHLHs(GlbHLHs)in ganod...Objective The basic helix-loop-helix(bHLH)transcription factors(TFs)are pivotal in regulating fungal growth,development,and secondary metabolism.However,the knowledge about the Ganoderma lucidum bHLHs(GlbHLHs)in ganoderic acid(GA)biosynthesis of G.lucidum was limited.This study aimed to explore the functions of bHLH genes in ganoderic acid biosynthesis during G.lucidum growth development.Methods First,the genome-wide identification of GlbHLHs was performed through Hidden Markov model searches and Two-way blast.Furthermore,through physicochemical properties,gene structure,and phylogenetic analysis,as well as combining the transcriptome and metabolome data from different developmental stages of G.lucidum,candidate GlbHLHs were screened.Subsequently,their regulatory roles in ganoderic acid biosynthesis were explored using yeast one-hybrid and dual-luciferase reporter assays.Results A total of 11 GlbHLH members were characterized in G.lucidum.The upstream promoter regions of these genes enriched hormones and abiotic stress responsive elements.Although individual ganoderic acid monomers demonstrated marked differences in accumulation patterns across specific growth phases and tissue types,overall,the total GA content was consistently higher in caps than in stipes throughout development.In addition,all GlbHLHs exhibited high expression in whole G.lucidum from the primordium to maturation stages.Among them,GlbHLH5 and GlbHLH7 showed the highest expression in any stage and highly correlated with key genes associated with GA pathway.Functional validation through dual-luciferase assays and yeast one-hybrid experiments had demonstrated that GlbHLH5 activated the P2 region of the lanosterol synthase promoter,while GlbHLH7 activated the promoters of squalene epoxidase and squalene synthase.Conclusion Compared to plants,G.lucidum harbored a small number of bHLH members but all high expression in any stages.Additionally,GlbHLH5 and GlbHLH7 with the highest expression among GlbHLHs showed activation in regulating the biosynthesis of GA.These results provide a theoretical reference for further research on ganoderic acid regulation in G.lucidum,and thereby providing a molecular foundation for enhancing ganoderic acid yield to optimize the medicinal value of G.lucidum.展开更多
The plant life cycle and the promise of crop yield start with successful seed germination,which requires an optimal balance between the phytohormones abscisic acid(ABA)and gibberellin(GA).Here,we report that the APETA...The plant life cycle and the promise of crop yield start with successful seed germination,which requires an optimal balance between the phytohormones abscisic acid(ABA)and gibberellin(GA).Here,we report that the APETALA 2-type transcription factor SALT AND ABA RESPONSE ERF 1(OsSAE1)antagonistically modulates ABA and GA signaling to control seed germination in rice(Oryza sativa L.).We show that knocking out OsSAE1 delays seed germination,concomitant with the accumulation of SLENDER RICE1(OsSLR1),a GA signaling repressor DELLA protein;importantly,GA application rescued the seed germination defect of ossae1 mutants.OsSAE1 directly activates transcription of the GA biosynthesis gene OsKS1 and represses that of the GA metabolism gene OsGA2ox3,resulting in higher GA levels.Moreover,OsSLR1physically interacts with ABA-INSENSITIVE 5(OsABI5),a key ABA signaling component,enhancing the transcriptional activation capacity of OsABI5 toward its target genes to regulate seed germination.The temporal expression pattern of OsSAE1 supports its role in orchestrating GA and ABA signaling to modulate seed germination and seed dormancy.Different OsSAE1 haplotypes differentially affected OsSAE1 transcript levels and seed germination rates,illustrating the potential of the elite OsSAE1 haplotype for genetic improvement of seed germination.Overall,our study reveals that OsSAE1 controls rice seed germination by regulating the balance between ABA and GA,providing a pivotal selection target for breeding rice cultivars suitable for direct seeding.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
Ferroptosis,a type of cell death that mainly involves iron metabolism imbalance and lipid peroxidation,is strongly correlated with the phagocytic response caused by bleeding after spinal cord injury.Thus,in this study...Ferroptosis,a type of cell death that mainly involves iron metabolism imbalance and lipid peroxidation,is strongly correlated with the phagocytic response caused by bleeding after spinal cord injury.Thus,in this study,bulk RNA sequencing data(GSE47681 and GSE5296)and single-cell RNA sequencing data(GSE162610)were acquired from gene expression databases.We then conducted differential analysis and immune infiltration analysis.Atf3 and Piezo1 were identified as key ferroptosis genes through random forest and least absolute shrinkage and selection operator algorithms.Further analysis of single-cell RNA sequencing data revealed a close relationship between ferroptosis and cell types such as macrophages/microglia and their intrinsic state transition processes.Differences in transcription factor regulation and intercellular communication networks were found in ferroptosis-related cells,confirming the high expression of Atf3 and Piezo1 in these cells.Molecular docking analysis confirmed that the proteins encoded by these genes can bind cycloheximide.In a mouse model of T8 spinal cord injury,low-dose cycloheximide treatment was found to improve neurological function,decrease levels of the pro-inflammatory cytokine inducible nitric oxide synthase,and increase levels of the anti-inflammatory cytokine arginase 1.Correspondingly,the expression of the ferroptosis-related gene Gpx4 increased in macrophages/microglia,while the expression of Acsl4 decreased.Our findings reveal the important role of ferroptosis in the treatment of spinal cord injury,identify the key cell types and genes involved in ferroptosis after spinal cord injury,and validate the efficacy of potential drug therapies,pointing to new directions in the treatment of spinal cord injury.展开更多
基金supported by the National Natural Science Foundation of China (81672028 and 81371788)。
文摘Human cytomegalovirus(HCMV)is a double-strand DNA virus widely infected in human.Circular RNAs(circ RNAs)are non-coding RNAs with most functions of which keep unknown,and the effects of HCMV productive infection on host circ RNA transcriptions remain unclear.In this study,we profiled 283 host circ RNAs that significantly altered by HCMV productive infection in human embryonic lung fibroblasts(HELF)by RNA deep sequencing and bioinformatics analysis.Among these,circ SP100,circ MAP3 K1,circ PLEKHM1,and circ TRIO were validated for their transcriptions and sequences.Furthermore,characteristics of circ SP100 were investigated by RT-q PCR and northern blot.It was implied that circ SP100 was produced from the sense strand of the SP100 gene containing six exons.Kinetics of circ SP100 and SP100 m RNA were significantly different after infection:circ SP100 levels increased gradually along with infection,whereas SP100 m RNA levels increased in the beginning and dropped at 24 h post-infection(hpi).Meanwhile,a total number of 257 proteins,including 10 HCMV encoding proteins,were identified potentially binding to cytoplasmic circ SP100 by RNA antisense purification(RAP)and mass spectrometry.Enrichment analysis showed these proteins were mainly involved in the spliceosome,protein processing,ribosome,and phagosome pathways,suggesting multiple functions of circ SP100 during HCMV infection.
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 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supported by National Key Research and Development Program of China(2022YFF1001600)BeijingNatural Science Foundation(5244040)+1 种基金STI2030-Major Projects(2030ZD0407101)China Postdoctoral Science Foundation(2022M723435)。
文摘Drought stress orchestrates a phosphorylation-dependent signaling cascade that reprograms transcriptional networks to enhance crop resilience.Through a large-scale transgenic screening,we identified ZmCRK5A,a Ca^(2+)-independent calcium-dependent protein kinase(CDPK)-related kinase,as a master regulator of drought tolerance in maize.Mechanistically,ZmCRK5A directly phosphorylates the MYB transcriptional repressor ZmSMH4(Single MYB Histone 4)at three conserved serine residues(Ser42/43/59)within its SANT domain,as demonstrated by in vitro kinase assays and site-directed mutagenesis.This post-translational modification abolishes Zm SMH4's DNA-binding capacity to ACC cis-elements,thereby de-repressing the potassium influx channel gene Zm KCH1(K^(+)Channel 1).Functional validation revealed that Zm KCH1 overexpression confers drought resilience through optimized stomatal dynamics and water retention,whereas clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-generated zmkch1 mutants display hypersensitivity to water deficit.Crucially,field evaluations demonstrated preserved grain yield alongside enhanced drought tolerance in plants with activated Zm CRK5A-Zm SMH4-Zm KCH1 signaling.Our findings delineate a kinase-transcription factor-ion channel axis that dynamically fine-tunes drought responses while maintaining productivity,providing a strategic framework for engineering stress-adapted crops without yield penalties.
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.
文摘Influenza viruses are a major cause of respiratory illness,with significant public health impact due to their ability to cause pandemics.This dialogue brought together experts including Professors George Fu Gao,Stephen Cusack,Mark von Itzstein,Ervin Fodor,Jonathan Grimes,Aartjan J.W.te Velthuis,and Tao Deng to decode the pressing scientific challenges and future directions in influenza research.They discussed how structural studies of the influenza polymerase have advanced our understanding of viral RNA transcription and replication.These insights are crucial for developing new antiviral drugs,with a particular focus on targeting the polymerase and its interactions with host factors like acidic nuclear phosphoprotein 32(ANP32).The dialogue also highlighted the potential of artificial intelligence(AI)to assist in designing small-molecule drugs,offering new strategies for combating influenza.Future research will continue to unravel the complexities of the polymerase’s role in replication,aiming to translate these findings into effective therapies and resilient public health strategies.
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.
文摘AIM:To investigate whether vaccinia-related kinase 1(VRK1)mediates transforming growth factor-beta2(TGF-β2)-caused epithelial-mesenchymal transition(EMT)and inflammatory responses in retinal pigment epithelial(RPE)cells through regulating snail family transcriptional repressor 1(SNAI1),and to validate its role in a proliferative vitreoretinopathy(PVR)mouse model.METHODS:Human RPE cell line ARPE-19 cells were treated with TGF-β2 to construct an EMT model.Western blot detected VRK1 level.The effects of VRK1 on SNAI1 expression and biological behavior of ARPE-19 cells were detected by immunofluorescence,ELISA,Transwell,and scratch assay,and the interaction between VRK1 and SNAI1 was confirmed through immunoprecipitation.A PVR mouse model was constructed,and the effects of VRK1 or/and SNAI1 on retinal damage were assessed by pathologic staining.Inflammatory factors and EMT-related proteins were assessed with ELISA and Western blot.RESULTS:VRK1 was upregulated in ARPE-19 cells after TGF-β2 treatment.Overexpression of VRK1 increased cell viability,promoted cell migration and EMT,and the levels of inflammatory factors.Silencing of VRK1 reversed the above indexes.There was a direct interaction between VRK1 and SNAI1,and overexpresssion SNAI1 weakened the impacts of silencing of VRK1.In PVR mice,silencing of VRK1 ameliorated retinal structural damage,decreased proinflammatory factor levels,and suppressed SNAI1 and mesenchymal marker expression.SNAI1 overexpression antagonized the protective effects of silencing VRK1 and exacerbated EMT and inflammatory responses.CONCLUSION:VRK1 plays a key role in retinal structural and inflammatory damage in PVR mice by regulating SNAI1 and mediating TGF-β2-caused EMT and inflammatory responses in RPE cells.
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.
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.
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.
基金funding from the Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(No.CI2021A04008)National Key Research and Development Project(No.2023YFC3504104)+2 种基金Hangzhou Joint Fund of the Zhejiang Provincial Natural Science Foundation of China(No.LHZSZ24H280003)Technology Major Program on Agricultural New Variety Breeding(No.2021C02073)Central Guiding Local Science and Technology Development Fund Project(No.2024ZY01009).
文摘Objective The basic helix-loop-helix(bHLH)transcription factors(TFs)are pivotal in regulating fungal growth,development,and secondary metabolism.However,the knowledge about the Ganoderma lucidum bHLHs(GlbHLHs)in ganoderic acid(GA)biosynthesis of G.lucidum was limited.This study aimed to explore the functions of bHLH genes in ganoderic acid biosynthesis during G.lucidum growth development.Methods First,the genome-wide identification of GlbHLHs was performed through Hidden Markov model searches and Two-way blast.Furthermore,through physicochemical properties,gene structure,and phylogenetic analysis,as well as combining the transcriptome and metabolome data from different developmental stages of G.lucidum,candidate GlbHLHs were screened.Subsequently,their regulatory roles in ganoderic acid biosynthesis were explored using yeast one-hybrid and dual-luciferase reporter assays.Results A total of 11 GlbHLH members were characterized in G.lucidum.The upstream promoter regions of these genes enriched hormones and abiotic stress responsive elements.Although individual ganoderic acid monomers demonstrated marked differences in accumulation patterns across specific growth phases and tissue types,overall,the total GA content was consistently higher in caps than in stipes throughout development.In addition,all GlbHLHs exhibited high expression in whole G.lucidum from the primordium to maturation stages.Among them,GlbHLH5 and GlbHLH7 showed the highest expression in any stage and highly correlated with key genes associated with GA pathway.Functional validation through dual-luciferase assays and yeast one-hybrid experiments had demonstrated that GlbHLH5 activated the P2 region of the lanosterol synthase promoter,while GlbHLH7 activated the promoters of squalene epoxidase and squalene synthase.Conclusion Compared to plants,G.lucidum harbored a small number of bHLH members but all high expression in any stages.Additionally,GlbHLH5 and GlbHLH7 with the highest expression among GlbHLHs showed activation in regulating the biosynthesis of GA.These results provide a theoretical reference for further research on ganoderic acid regulation in G.lucidum,and thereby providing a molecular foundation for enhancing ganoderic acid yield to optimize the medicinal value of G.lucidum.
基金funded by the National Natural Science Foundation of China(32472037,32030079)the National Key Research and Development Program of China(2022YFD1201700)+2 种基金the Youth innovation of Chinese Academy of Agricultural Sciences(Y2024QC14)the Central Public-Interest Scientific Institution Basal Research Fund(1610392023004)Agricultural Science and Technology Innovation Program(CAAS-ZDRW202407)。
文摘The plant life cycle and the promise of crop yield start with successful seed germination,which requires an optimal balance between the phytohormones abscisic acid(ABA)and gibberellin(GA).Here,we report that the APETALA 2-type transcription factor SALT AND ABA RESPONSE ERF 1(OsSAE1)antagonistically modulates ABA and GA signaling to control seed germination in rice(Oryza sativa L.).We show that knocking out OsSAE1 delays seed germination,concomitant with the accumulation of SLENDER RICE1(OsSLR1),a GA signaling repressor DELLA protein;importantly,GA application rescued the seed germination defect of ossae1 mutants.OsSAE1 directly activates transcription of the GA biosynthesis gene OsKS1 and represses that of the GA metabolism gene OsGA2ox3,resulting in higher GA levels.Moreover,OsSLR1physically interacts with ABA-INSENSITIVE 5(OsABI5),a key ABA signaling component,enhancing the transcriptional activation capacity of OsABI5 toward its target genes to regulate seed germination.The temporal expression pattern of OsSAE1 supports its role in orchestrating GA and ABA signaling to modulate seed germination and seed dormancy.Different OsSAE1 haplotypes differentially affected OsSAE1 transcript levels and seed germination rates,illustrating the potential of the elite OsSAE1 haplotype for genetic improvement of seed germination.Overall,our study reveals that OsSAE1 controls rice seed germination by regulating the balance between ABA and GA,providing a pivotal selection target for breeding rice cultivars suitable for direct seeding.
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.
基金supported by the National Natural Science Foundation of China,No.81972073(to HZ)a grant from the Taishan Scholars Program ofShandong Province-Young Taishan Scholars,No.tsqn201909197(to HZ)+1 种基金a grant from Tianjin Key Medical Discipline(Specialty)Construct Project,No.TJYXZDXK-027A(to SF)a grant from Academic Expert International Innovation Summit,No.22JRRCRC00010(to SF).
文摘Ferroptosis,a type of cell death that mainly involves iron metabolism imbalance and lipid peroxidation,is strongly correlated with the phagocytic response caused by bleeding after spinal cord injury.Thus,in this study,bulk RNA sequencing data(GSE47681 and GSE5296)and single-cell RNA sequencing data(GSE162610)were acquired from gene expression databases.We then conducted differential analysis and immune infiltration analysis.Atf3 and Piezo1 were identified as key ferroptosis genes through random forest and least absolute shrinkage and selection operator algorithms.Further analysis of single-cell RNA sequencing data revealed a close relationship between ferroptosis and cell types such as macrophages/microglia and their intrinsic state transition processes.Differences in transcription factor regulation and intercellular communication networks were found in ferroptosis-related cells,confirming the high expression of Atf3 and Piezo1 in these cells.Molecular docking analysis confirmed that the proteins encoded by these genes can bind cycloheximide.In a mouse model of T8 spinal cord injury,low-dose cycloheximide treatment was found to improve neurological function,decrease levels of the pro-inflammatory cytokine inducible nitric oxide synthase,and increase levels of the anti-inflammatory cytokine arginase 1.Correspondingly,the expression of the ferroptosis-related gene Gpx4 increased in macrophages/microglia,while the expression of Acsl4 decreased.Our findings reveal the important role of ferroptosis in the treatment of spinal cord injury,identify the key cell types and genes involved in ferroptosis after spinal cord injury,and validate the efficacy of potential drug therapies,pointing to new directions in the treatment of spinal cord injury.
