To develop herbicides with a novel mechanism of action,a series of 1,3,4-oxadiazolpyridine derivatives were designed and synthesized based on active substructure splicing and structure optimization.These derivatives(5...To develop herbicides with a novel mechanism of action,a series of 1,3,4-oxadiazolpyridine derivatives were designed and synthesized based on active substructure splicing and structure optimization.These derivatives(5aa-5bd)were characterized by their melting points,^(1)H and^(13)C nuclear magnetic resonance spectroscopy,and high-resolution mass spectrometry.The configuration of 5 al was determined using single-crystal X-ray diffraction.Additionally,5 al exhibited excellent herbicidal activity at a dosage of 75 g/hm^(2),showing an EC 50 of 4.03 g/hm^(2)against both E.crus-galli and quinclorac-resistant E.crus-galli.At a dosage of 375 g/hm 2,5 al was safe for application on rice and sorghum and showed low toxicity(>200μg/g)towards Apis mellifera.After treatment with 5 al,the lamellae of the chloroplast grana of barnyard grass leaves were stacked disorderly and arranged loosely,and some thylakoids were broken,as observed by transmission electron microscopy.Transcriptomics analysis of E.crus-galli revealed that 5 al affects the defense response,membranes,plasma membranes,and chloroplasts of differentially expressed genes,which alter membrane permeability and energy metabolism,potentially leading to plant death.Thus,we successfully developed a novel molecular scaffold with a new mechanism of action that exhibits herbicidal activity against resistant E.crus-galli.Therefore,further development of lead herbicides based on this scaffold is required.展开更多
The formation of root system architecture(RSA)plays a crucial role in plant growth.OsDRO1 is known to have a function in controlling RSA in rice,however,the role of potato StDRO2,a homolog of rice OsDRO1,in root growt...The formation of root system architecture(RSA)plays a crucial role in plant growth.OsDRO1 is known to have a function in controlling RSA in rice,however,the role of potato StDRO2,a homolog of rice OsDRO1,in root growth remains unclear.In this study,we obtained potato dro2 mutant lines by Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-Associated 9(CRISPR/Cas9)-mediated genome editing system.The mutant lines were generated from a splicing defect of the StDRO2 intron 1,which causes a nonsense mutation in StDRO2.Furthermore,the secondary structure of StDRO2 mRNA analyzed with RNAfold Web Server was altered in the dro2 mutant.Mutation of StDRO2 conveys potato adaptation through changing the RSA via alteration of auxin transport under drought stress.The potato dro2 lines showed higher plant height,longer root length,smaller root growth angle and increased tuber weight than the wild-type.The alteration of RSA was associated with a disturbance of IAA distribution in the dro2 mutant,and the levels of StPIN7 and StPIN10 detected by using real-time PCR were up-regulated in the roots of potato dro2 lines grown under drought stress.Moreover,the microRNAs(miRNAs)PmiREN024536 and PmiREN024486 targeted the StDRO2 gene,and auxin positively and negatively regulated the expression of StDRO2 and the miRNAs PmiREN024536 and PmiREN024486,respectively,in the potato roots.Our data shows that a regulatory network involving auxin,StDRO2,PmiREN024536 and PmiREN024486 can control RSA to convey potato fitness under drought stress.展开更多
The steel tube arch rib in a large-span concrete-filled steel tube arch bridge has a large span and diameter,which also leads to a larger weld seam scale.Large-scale welding seams will inevitably cause more obvious we...The steel tube arch rib in a large-span concrete-filled steel tube arch bridge has a large span and diameter,which also leads to a larger weld seam scale.Large-scale welding seams will inevitably cause more obvious welding residual stress(WRS).For the purpose of studying the influence of WRS from large-scale welding seam on the mechanical properties of steel tube arch rib during arch rib splicing,test research and numerical simulation analysis on the WRS in arch rib splicing based on the Guangxi Pingnan Third Bridge,which is the world’s largest span concrete-filled steel tube arch bridge,were conducted in this paper,and the distribution pattern of WRS at the arch rib splicing joint was obtained.Subsequently,the WRS was introduced into the mechanical performance analysis of joints and structures to analyze its effects.The findings reveal that the distribution of WRS in the arch rib is greatly influenced by the rib plate,and the axial WRS in the heat-affected zone are primarily tensile,while the circumferential WRS are distributed in an alternating pattern of tensile and compressive stresses along the circumferential direction of the main tube.Under the influence of WRS,the ultimate bearing capacity of the joint is reduced by 29.4%,the initial axial stiffness is reduced by 4.32%,and the vertical deformation of the arch rib structure is increased by 4.7%.展开更多
Objectives:KH-type splicing regulatory protein(KHSRP)is an RNA-binding protein involved in several cellular processes,including nuclear splicing,mRNA localization,and cytoplasmic degradation.While KHSRP’s role has be...Objectives:KH-type splicing regulatory protein(KHSRP)is an RNA-binding protein involved in several cellular processes,including nuclear splicing,mRNA localization,and cytoplasmic degradation.While KHSRP’s role has been studied in other cancers,its specific involvement in gastric cancer remains poorly understood.This study aims to explore KHSRP expression in gastric cancer and its potential effects on tumor progression and immune response.Methods:KHSRP expression in gastric cancer tissues and normal tissues was analyzed using data from The Cancer Genome Atlas(TCGA)database.The correlation between KHSRP expression,patient survival,and immune response was also assessed.Immunohistochemistry was performed to evaluate KHSRP expression in gastric cancer tissues.Gain-and loss-of-function experiments were conducted to assess KHSRP’s effects on gastric cancer cell proliferation,stemness,and migration.Furthermore,the impact of KHSRP silencing on tumor volume and immune cell infiltration was evaluated in a C3H/He mouse xenograft model.Results:KHSRP was found to be overexpressed in gastric cancer tissues compared to normal tissues,with a positive correlation to tumor stage and a negative correlation with patient prognosis.Functional assays revealed that KHSRP promotes gastric cancer cell proliferation,enhances cancer stem cell properties,and increases migratory capabilities in vitro.In vivo,KHSRP silencing led to a significant reduction in tumor volume and increased immune cell infiltration in the mouse xenograft model.Conclusions:KHSRP acts as an oncogene in gastric cancer by promoting tumorigenesis and suppressing anti-tumor immune responses.Its overexpression is associated with poor prognosis,making KHSRP a potential prognostic marker and therapeutic target in gastric cancer.展开更多
Pentatricopeptide repeat(PPR)proteins are a large group of eukaryote-specific RNA-binding proteins that play pivotal roles in plant organelle gene expression.Here,we report the function of PPR21 in mitochondrial intro...Pentatricopeptide repeat(PPR)proteins are a large group of eukaryote-specific RNA-binding proteins that play pivotal roles in plant organelle gene expression.Here,we report the function of PPR21 in mitochondrial intron splicing and its role in maize kernel development.PPR21 is a typical P-type PPR protein targeted to mitochondria.The ppr21 mutants are arrested in embryogenesis and endosperm development,leading to embryo lethality.Null mutations of PPR21 reduce the splicing efficiency of nad2 intron 1,2,and 4 and impair the assembly and activity of mitochondrial complex I.Previous studies show that the P-type PPR protein EMP12 is required for the splicing of identical introns.However,our protein interaction analyses reveal that PPR21 does not interact with EMP12.Instead,both PPR21 and EMP12 interact with the small MutS-related(SMR)domain-containing PPR protein 1(PPR-SMR1)and the short P-type PPR protein 2(SPR2).PPR-SMR1 interacts with SPR2,and both proteins are required for the splicing of many introns in mitochondria,including nad2 intron 1,2,and 4.These results suggest that a PPR21-(PPR-SMR1/SPR2)-EMP12 complex is involved in the splicing of nad2 introns in maize mitochondria.展开更多
In this study,we examine the problem of sliced inverse regression(SIR),a widely used method for sufficient dimension reduction(SDR).It was designed to find reduced-dimensional versions of multivariate predictors by re...In this study,we examine the problem of sliced inverse regression(SIR),a widely used method for sufficient dimension reduction(SDR).It was designed to find reduced-dimensional versions of multivariate predictors by replacing them with a minimally adequate collection of their linear combinations without loss of information.Recently,regularization methods have been proposed in SIR to incorporate a sparse structure of predictors for better interpretability.However,existing methods consider convex relaxation to bypass the sparsity constraint,which may not lead to the best subset,and particularly tends to include irrelevant variables when predictors are correlated.In this study,we approach sparse SIR as a nonconvex optimization problem and directly tackle the sparsity constraint by establishing the optimal conditions and iteratively solving them by means of the splicing technique.Without employing convex relaxation on the sparsity constraint and the orthogonal constraint,our algorithm exhibits superior empirical merits,as evidenced by extensive numerical studies.Computationally,our algorithm is much faster than the relaxed approach for the natural sparse SIR estimator.Statistically,our algorithm surpasses existing methods in terms of accuracy for central subspace estimation and best subset selection and sustains high performance even with correlated predictors.展开更多
BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on th...BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on the evaluation of the potential role of PSMD6 in hepatocellular carcinoma(HCC).AIM To comprehensively evaluate the overexpression pattern and clinical significance of PSMD6 in HCC tissues.METHODS This study integrated PSMD6 mRNA expression profiles from 4672 HCC and 3667 non-HCC tissues,along with immunohistochemical scores from 383 HCC and adjacent tissues,to assess PSMD6 overexpression in HCC.Clustered regularly interspaced short palindromic repeats knockout technology evaluated PSMD6’s essential role in HCC cell growth.Functional enrichment analysis explored the molecular mechanism of PSMD6 abnormalities in HCC.Drug sensitivity analysis and molecular docking analysed the effect of abnormal expression of PSMD6 on the drug sensitivity of HCC cells.RESULTS The results of 41 external and two internal datasets showed that PSMD6 mRNA(SMD=0.26,95%CI:0.09-0.42,P<0.05)and protein(SMD=2.85,95%CI:1.19-4.50,P<0.05)were significantly overexpressed in HCC tissues.The integrated analysis results showed that PSMD6 had a significant overexpression pattern in HCC tissues(SMD=0.40,95%CI:0.15-0.66,P<0.05).PSMD6 knockout inhibited HCC cell growth(chronos scores<-1).Functional enrichment implicated ribosome biogenesis and RNA splicing.Significant enrichment of signalling pathways such as RNA degradation,ribosomes,and chemical carcinogenesis—reactive oxygen species.Drug sensitivity analysis and a molecular docking model showed that high expression of PSMD6 was associated with the tolerance of HCC cells to drugs such as ML323,sepantronium bromide,and GDC0810.Overexpressed PSMD6 effectively distinguished HCC tissues(AUC=0.75,95%CI:0.71-0.79).CONCLUSION This study was the first to discover that PSMD6 was overexpressed in HCC tissues.PSMD6 is essential for the growth of HCC cells and may be involved in ribosome biogenesis and RNA splicing.展开更多
Stimulus-responsive actuators are novel functional devices capable of sensing external stimuli and ex-hibiting specific deformation responses.MXene,owing to its unique 2D structure and efficient energy conversion effi...Stimulus-responsive actuators are novel functional devices capable of sensing external stimuli and ex-hibiting specific deformation responses.MXene,owing to its unique 2D structure and efficient energy conversion efficiency,has bridged the gap in traditional devices and shown great potential for multiple stimulus-responsive actuators.However,the drawbacks of pure MXene films,including susceptibility to oxidation and vulnerability to shear stress,hinder their applications.Through composite modification and structural design strategies,a three-layer structured MXene-carbon nanotubes hybrid film(tHCM)is fabri-cated,exhibiting a tensile strength and fracture strain of 153.8 MPa and 4.65%,respectively,representing improvements of 598.4%and 226.8%compared to the initial film.Meanwhile,the film maintains excel-lent stability demonstrating the enhancing effects of hydrogen bonds and densely packed structure.The hybrid films demonstrate unique and facile welding features due to splicing properties,enabling the for-mation of complex configurations.In terms of electro-/photo-thermal conversion performance,the hybrid film can reach a reasonably high temperature of 250℃at low voltage(2.5 V)and 110.6℃under 150 mW cm^(-2) infrared light.Leveraging the thermal expansion mismatch between tHCM and thermoplastic films,an integrated,flexible,and weldable actuator with unique electro/photo-response is developed,and vari-ous biomimetic driving applications,particularly,the light-mediated hierarchical transmission and precise motion along predetermined trajectory are realized.This work not only provides an effective strategy for modifying MXene composite films but also advances the design of novel actuators,offering broad appli-cation prospects in fields such as stimulus-responsive actuated robots and cargo transportation.展开更多
Caspases,which play key roles in cell apoptosis,undergo alternative splicing to form different splicing variants that can regulate the apoptotic process.Lepidopteran insect caspases undergo alternative splicing,althou...Caspases,which play key roles in cell apoptosis,undergo alternative splicing to form different splicing variants that can regulate the apoptotic process.Lepidopteran insect caspases undergo alternative splicing,although the functions of their splicing variants are still unclear.The Spodoptera exigua caspase-5(SeCaspase-5)gene was cloned and found to produce four different splicing variants with different gene sequences and protein functional domains,which were named SeCaspase-5a,SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Overexpression of these variants in S.exigua cells(Se-3)showed that SeCaspase-5a had a proapoptotic function,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not.Semi-qPCR analysis revealed that the expression of the SeCaspase-5 variants significantly differed during Autographa californica multiple nucleopolyhedrovirus(AcMNPV)infection.Furthermore,the SeCaspase-5 variants were constructed into the AcMNPV bacmid and transfected into Se-3 cells,which revealed that SeCaspase-5a promoted cell apoptosis and reduced virus production,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not promote cell apoptosis but instead increased virus production.Moreover,an analysis of the interactions between the SeCaspase-5 variants revealed that SeCaspase-5a directly interacted with SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Coexpression of these variants in Se-3 cells also revealed that SeCaspase-5b,SeCaspase-5c and SeCaspase-5d inhibited the proapoptotic function of SeCaspase-5a,resulting in a reduction in the percentage of apoptotic cells by about 20%.These results indicate that SeCaspase-5 undergoes alternative splicing and is involved in regulating the apoptosis induced by baculovirus infection.These findings increase our understanding of the functions of lepidopteran insect caspases and provide new insights into the mechanism of host-cell apoptosis induced by baculoviruses.展开更多
Survival of motor neuron(SMN)protein encoded by SMN1 gene,is the essential and ubiquitously expressed protein in all tissues.Prior studies demonstrated that SMN deficiency impaired bone development,but the underlying ...Survival of motor neuron(SMN)protein encoded by SMN1 gene,is the essential and ubiquitously expressed protein in all tissues.Prior studies demonstrated that SMN deficiency impaired bone development,but the underlying mechanism of abnormal endochondral ossification remains obscure.Here,we showed SMN is involved in hypertrophic chondrocytes differentiation through regulating RNA splicing and protein degradation via analyzing single cell RNA-sequencing data of hypertrophic chondrocytes.Of note,SMN loss induced dwarfism and delayed endochondral ossification in Smn1 depletion-severe spinal muscular atrophy(SMA)mouse model and Smn1 chondrocyte conditional knockdown mouse.Histological analysis revealed that SMN deficiency expanded the zone of hypertrophic chondrocytes in the growth plates,but delayed turnover from hypertrophic to ossification zone.Widespread changes in endochondral ossification related gene expression and alternative splicing profiles were identified via RNA sequencing of growth plate cartilages from SMA mice on postnatal day 4.Importantly,Mass spectrometry-based proteomics analysis elucidated Y-box-binding protein 1(YBX1)as a vital SMN-binding factor,was decreased in SMA mice.YBX1 knockdown reproduced the aberrant gene expression and splicing changes observed in SMA growth plate cartilages.Comparing the binding proteins of SMN and YBX1 revealed TNF receptor-associated factor 6(TRAF6),which promoted ubiquitination degradation of YBX1.By conditionally deleting Smn1 in chondrocytes of WT mice and overexpressing Smn1 in chondrocytes of SMA mice,we proved that SMN expression in chondrocytes is critical for hypertrophic chondrocyte-mediated endochondral ossification.Collectively,these results demonstrate that SMN deficiency contributes to rapid systemic bone dysplasia syndrome by promoting TRAF6-induced ubiquitination degradation of YBX1 in growth plate cartilages of SMA mice.展开更多
Male infertility constitutes a major global public health concern,with the underlying etiology remaining unidentified in nearly half of the diagnosed cases.Protein kinase CK1α(CK1α)functions as a pivotal regulator o...Male infertility constitutes a major global public health concern,with the underlying etiology remaining unidentified in nearly half of the diagnosed cases.Protein kinase CK1α(CK1α)functions as a pivotal regulator of cell cycle progression,pre-mRNA processing,and spliceosome-associated pathways through interactions with specific splicing factors.Comprehensive analyses revealed CK1αexpression in both germ cells and somatic cells of mouse testes,implicating its involvement in spermatogenic regulation.However,the physiological roles and mechanistic basis of CK1αfunction in Sertoli cells remain unclear.In this study,CK1αwas highly expressed in Sertoli cells,and conditional knockout of CK1αin murine Sertoli cells induced profound testicular atrophy and complete infertility.This phenotype was characterized by rapid depletion of Sertoli cells and spermatogenic dysfunction.Subsequent analyses demonstrated that CK1αregulated the fate determination of fetal and neonatal Sertoli cells in mice.At the molecular level,CK1αpromoted Sertoli cell survival through interaction with the splicing factor ZRSR1 to modulate apoptosis.Collectively,these findings establish CK1αas a key regulator of alternative splicing and male reproduction,providing critical insights into the molecular mechanisms underlying testicular development and reproductive function.展开更多
ObjectivesThe PTPRQ gene is essential for preserving the structure and function of stereocilia in inner ear.However,research on splicing mutations within this gene is limited.This study aims to investigate novel splic...ObjectivesThe PTPRQ gene is essential for preserving the structure and function of stereocilia in inner ear.However,research on splicing mutations within this gene is limited.This study aims to investigate novel splicing mutations in PTPRQ,clarify their molecular mechanisms,and provide new insights into the genetic factors associated with hearing loss,ultimately enhancing diagnostic accuracy.MethodClinical data and peripheral blood samples were obtained from members of a family with congenital hearing loss.Variants were identified through high-throughput sequencing and confirmed by Sanger sequencing to ensure genealogical co-segregation.The splicing effects of PTPRQ variants were evaluated using bioinformatics tools and minigene assays.ResultsWe used whole exome sequencing to identify novel double compound heterozygous splice-altering variants(c.5426+1 G>A and c.6603-3 T>G)in the PTPRQ gene with DFNB84A.We molecularly characterized these variants,and they were found to co-segregate with the disease within the family.Minigene assays and Sanger sequencing confirmed that the c.6603-3 T>G variant caused exon 43 skipping,resulting in a frameshift mutation(p.Ser2201ArgfsTer112).Further bioinformatic analysis supported these findings.ConclusionsThis study identifies a novel compound heterozygous splicing variant in the PTPRQ gene in a Chinese family with DFNB84A,expanding the known spectrum of PTPRQ mutations.These findings enhance the understanding of PTPRQ-related hearing loss and may aid in early diagnosis,prevention,and therapeutic strategies.展开更多
The plant circadian clock temporally drives gene expression throughout the day and coordinates various physiological processes with diurnal environmental changes. It is essential for conferring plant fitness and compe...The plant circadian clock temporally drives gene expression throughout the day and coordinates various physiological processes with diurnal environmental changes. It is essential for conferring plant fitness and competitive advantages to survive and thrive under natural conditions through the circadian control of gene transcription. Chinese cabbage(Brassica rapa ssp. pekinensis) is an economically important vegetable crop worldwide, although there is little information concerning its circadian clock system. Here we found that gene expression patterns are affected bycircadian oscillators at both the transcriptional and post-transcriptional levels in Chinese cabbage. Time-course RNA-seq analyses were conducted on two short-period lines(SPcc-1 and SPcc-2) and two long-period lines(LPcc-1 and LPcc-2) under constant light. The results showed that 32.7–50.5% of the genes were regulated bythe circadian oscillator and the expression peaks of cycling genes appeared earlier in short-period lines than long-period lines. In addition, approximately 250 splicing events exhibited circadian regulation, with intron retention(IR) accounting for a large proportion. Rhythmically spliced genes included the clock genes LATE ELONGATEDHYPOCOTYL(BrLHY), REVEILLE 2(BrRVE2) and EARLY FLOWERING 3(BrELF3). We also found that thecircadian oscillator could notably influence the diurnal expression patterns of genes that are associated with glucose metabolism via photosynthesis, the Calvin cycle and the tricarboxylic acid(TCA) cycle at both the transcriptional andpost-transcriptional levels. The collective results of this study demonstrate that circadian-regulated physiological processes contribute to Chinese cabbage growth and development.展开更多
Background:Alterations in splicing factors contribute to aberrant alternative splicing(AS),which subsequently promotes tumor progression.The splicing factor polypyrimidine tract binding protein 1(PTBP1)has been shown ...Background:Alterations in splicing factors contribute to aberrant alternative splicing(AS),which subsequently promotes tumor progression.The splicing factor polypyrimidine tract binding protein 1(PTBP1)has been shown to facilitate cancer progression by modulating oncogenic variants.However,its specific role and underlying mechanisms in hepatocellular carcinoma(HCC)remain to be elucidated.Methods:PTBP1 expression was evaluated in HCC tissues and cell lines.Subsequently,cells were transfected with vectors designed for PTBP1 overexpression or downregulation.The biological function of PTBP1 was assessed in vitro and in vivo using MTS assays,colony formation assays,transwell assays,xenograft formation,tail vein injection,and orthotopic models.Transcriptome analysis was conducted to elucidate the underlying molecular mechanisms.Results:Our findings demonstrated that PTBP1 exhibited elevated expression in HCC cell lines and tissues.Furthermore,its expression positively correlated with overall and disease-free survival rates,as well as tumor grade and stage.PTBP1 knockdown reduced HCC cell proliferation,migration,and invasion in vitro and suppressed hepatocarcinoma xenograft growth and infiltration in vivo.RNA sequencing(RNA-Seq)analysis identified the AS events associated with PTBP1.PTBP1 functionally enhanced cell proliferation,invasion,and migration by modulating the AS of the microtubule-associated protein tau(MAPT)gene and promoting oncogene expression.Notably,the dysregulation of MAPT splicing coincided with increased PTBP1 expression in HCC.Conclusions:PTBP1-guided AS of the MAPT gene enhances tumorigenicity in HCC through activation of the MAPK/ERK pathways.展开更多
Heat stress is a major threat to maize(Zea mays L.)production worldwide.Heat shock transcription factors(HSFs)play vital roles in plant responses to heat stress.However,the molecular and genetic mechanisms underlying ...Heat stress is a major threat to maize(Zea mays L.)production worldwide.Heat shock transcription factors(HSFs)play vital roles in plant responses to heat stress.However,the molecular and genetic mechanisms underlying HSF-meditated thermotolerance in maize remain largely unexplored.In this study,we demonstrate that the alternative splicing of ZmHsf23 modulates heat stress tolerance in maize.Hsf23 produced two functional transcripts,Hsf23L and Hsf23S,which differ by the presence of a cryptic mini-exon in Hsf23L that is spliced out in Hsf23S.Both transcripts were strongly induced by heat stress.Mutants lacking Hsf23L alone(hsf23l)or both Hsf23L and Hsf23S(hsf23l23s)exhibited increased susceptibility to heat stress,whereas overexpression of Hsf23S enhanced heat stress tolerance in maize.Subsequently,we found that Hsf23S positively regulates heat stress tolerance by directly activating the transcription of three sHSP genes(Hsp16.9,Hsp17.2,and Hsp18a)and TIL1 gene.In addition,Hsf23L physically interacted with Hsf23S and enhanced the transcriptional activation of Hsf23S on the sHSPs and TIL1 promoters.Notably,genetic analysis suggested that co-overexpression of Hsf23L and Hsf23S further improves heat tolerance of the transgenic plants.Taken together,these results reveal two splicing variants of ZmHsf23 cooperatively regulate maize heat tolerance,thus highlighting potential value of ZmHsf23 in breeding heat-tolerant maize varieties.展开更多
Starch biosynthesis is a complex process that relies on the coordinated action of multiple enzymes.Resistant starch is not digested in the small intestine,thus preventing a rapid rise in the glycemic index.Starch synt...Starch biosynthesis is a complex process that relies on the coordinated action of multiple enzymes.Resistant starch is not digested in the small intestine,thus preventing a rapid rise in the glycemic index.Starch synthase 2a(SS2a)is a key enzyme in amylopectin biosynthesis that has significant effects on starch structure and properties.In this study,we identified an ss2a null mutant(M3-1413)with a single base mutation from an ethyl methane sulfonate(EMS)-mutagenized population of barley.The mutation was located at the 3'end of the first intron of the RNA splicing receptor(AG)site,and resulted in abnormal RNA splicing and two abnormal transcripts of ss2a,which caused the inactivation of the SS2a gene.The starch structure and properties were significantly altered in the mutant,with M3-1413 containing lower total starch and higher amylose and resistant starch levels.This study sheds light on the effect of barley ss2a null mutations on starch properties and will help to guide new applications of barley starch in the development of nutritious food products.展开更多
The silver-lipped pearl oyster(Pinctada maxima)is the largest and most commercially valuable pearl-producing oyster,renowned for its ability to generate large,lustrous pearls.This species is a sequential hermaphrodite...The silver-lipped pearl oyster(Pinctada maxima)is the largest and most commercially valuable pearl-producing oyster,renowned for its ability to generate large,lustrous pearls.This species is a sequential hermaphrodite,with pearl production displaying notable sexual dimorphism.Consequently,understanding the molecular mechanisms governing sex determination and differentiation is crucial for advancing breeding strategies in the pearl oyster industry.To elucidate these mechanisms,this study conducted integrative transcriptomic analyses of P.maxima gonadal tissues using isoform sequencing(Isoseq)and RNA sequencing(RNA-seq).Comparative analysis of ovarian and testicular tissues identified 2768 differentially expressed genes(DEGs).Gene coexpression network analysis delineated four key modules,including three sex-specific modules and one shared module.Key genes implicated in sex determination and maintenance were identified,including FOXL2,NANOS1,andβ-catenin,important for ovarian maintenance,and DMRT,SOX30,FEM1,and FOXJ1,crucial for testicular maintenance.These genes,widely studied in other taxa,were confirmed as hub genes in the sex-related modules of P.maxima.Interestingly,genes within the shared module were significantly enriched in the spliceosome pathway.Alternative splicing analysis highlighted its extensive role in gonadal tissues,with more pronounced activity observed in the testis compared to the ovary.Nearly half(47.83%,375)of the identified genes undergoing differential alternative splicing(DASGs)also exhibited differential transcript usage(DTUGs),while only 17%of DTUGs overlapped with DEGs.Genes associated with sex differentiation,such as DMRT,β-catenin,and U2AF2,displayed sex-specific and/or sex-biased isoforms.These findings offer novel insights into the molecular basis of sex differentiation in P.maxima,which could inform the development of targeted breeding strategies aimed at sex control,thereby enhancing pearl quality and yield in aquaculture.This study offers a robust molecular foundation for advancing breeding programs and optimizing production in the pearl oyster industry.展开更多
Background:Glioblastoma(GBM)is one of the most malignant types of central nervous system tumors.Oxygen deprivation in the tumor microenvironment is thought to be an important factor in promoting GBM progression.Howeve...Background:Glioblastoma(GBM)is one of the most malignant types of central nervous system tumors.Oxygen deprivation in the tumor microenvironment is thought to be an important factor in promoting GBM progression.However,the mechanisms of hypoxia-promoted tumor progression remain elusive.Methods:Alternative splicing of diacylglycerol kinase gamma(DGKG)-Δexon13 was amplified and verified by PCR-Sanger sequencing.The functions of DGKG and DGKG-Δexon13 were analyzed by Cell counting kit-8(CCK-8),Transwell,Matrigeltranswell experiments,and in vivo orthotropic GBM animal models.Transcriptome analyses were done to find out the regulated genes.Results:In this study,we found that a new transcript DGKG-Δexon13 was generated in GBM under hypoxia via alternative splicing.Moreover,the results of CCK-8,Transwell,and Matrigel-transwell experiments showed that the proliferation,migration,and invasion abilities of U87-MG and T98G were decreased after DGKG knockdown.Compared to wild-type DGKG,DGKG-Δexon13 overexpression significantly promoted cellular proliferation,migration,and invasion abilities in GBM.Furthermore,in vivo,orthotropic GBM animal models analysis showed that the tumor volumes were much smaller in the DGKG knockdown group.However,the tumor sizes in the DGKG and DGKG-Δexon13 rescue groups were restored,especially in the DGKG-Δexon13 group.Transcriptome analysis revealed that MORC1,KLHDC7B,ATP1A2,INHBE,TMEM119,and FGD3 were altered significantly when DGKG was knocked down.IL-16,CCN2,and EFNB3 were specifically regulated by DGKG-Δexon13.Conclusions:Our study found that hypoxia-induced alternative splicing transcript DGKG-Δexon13 promotes GBM proliferation and infiltration,which might provide a new potential target for the clinical treatment and diagnosis of GBM.展开更多
To address the installation challenges of a 2-m ring Gregorian telescope system,and similar optical systems with a small width-to-radius ratio,we propose a detection method combining local interferometry with a compar...To address the installation challenges of a 2-m ring Gregorian telescope system,and similar optical systems with a small width-to-radius ratio,we propose a detection method combining local interferometry with a comparison model.This method enhances the precision of system calibration by establishing a dataset that delineates the relationship between secondary mirror misalignment and wavefront aberration,subsequently inferring the misalignment from interferometric detection results during the calibration process.For the 2-m ring telescope,we develop a detection model using five local sub-apertures,enabling a root-mean-square detection accuracy of 0:0225λ(λ=632:8 nm)for full-aperture wavefront aberration.The calibration results for the 2-m Ring Solar Telescope system indicate that the root-mean-square value of sub-aperture wavefront aberration reaches 0.104λ,and the root-mean-square value of spliced full-aperture measurement yields reaches 0.112λ.This method offers a novel approach for calibrating small width-toradius ratio telescope systems and can be applied to the calibration of other irregular-aperture optical systems.展开更多
基金supported by the National Key Research and Development Program of China(2023YFD1400504)Natural Science Foundation of Hunan Province(2024JJ2036)+3 种基金Natural Science Foundation of China(32172433)Foundation for Tobacco Science of China National Tobacco Corporation(110202401015,(LS-05))Scientific-Innovative of Hunan Agricultural Sciences and Technology(2024CX69)China Agriculture Research System of MOF and MARA(CARS-16-E19)。
文摘To develop herbicides with a novel mechanism of action,a series of 1,3,4-oxadiazolpyridine derivatives were designed and synthesized based on active substructure splicing and structure optimization.These derivatives(5aa-5bd)were characterized by their melting points,^(1)H and^(13)C nuclear magnetic resonance spectroscopy,and high-resolution mass spectrometry.The configuration of 5 al was determined using single-crystal X-ray diffraction.Additionally,5 al exhibited excellent herbicidal activity at a dosage of 75 g/hm^(2),showing an EC 50 of 4.03 g/hm^(2)against both E.crus-galli and quinclorac-resistant E.crus-galli.At a dosage of 375 g/hm 2,5 al was safe for application on rice and sorghum and showed low toxicity(>200μg/g)towards Apis mellifera.After treatment with 5 al,the lamellae of the chloroplast grana of barnyard grass leaves were stacked disorderly and arranged loosely,and some thylakoids were broken,as observed by transmission electron microscopy.Transcriptomics analysis of E.crus-galli revealed that 5 al affects the defense response,membranes,plasma membranes,and chloroplasts of differentially expressed genes,which alter membrane permeability and energy metabolism,potentially leading to plant death.Thus,we successfully developed a novel molecular scaffold with a new mechanism of action that exhibits herbicidal activity against resistant E.crus-galli.Therefore,further development of lead herbicides based on this scaffold is required.
基金supported by grants from the National Natural Science Foundation of China(Grant Nos.32260085,31860064,31660501,31970609,32260718 and 31901870)the Key Projects of the Applied Basic Research Plan of Yunnan Province(Grant No.202301AS070082)+3 种基金the Start-up fund from Xishuangbanna Tropical Botanical Garden,the‘Top Talents Program in Science and Technology’from Yunnan Province,the Major Science and Technology Project in Yunnan Province(Grant Nos.202102AE090042 and 202202AE090036)the Young and Middle-Aged Academic and Technical Leaders Reserve Talent Program in Yunnan Province(Grant No.202205AC160076)China Postdoctoral Science Foundation(Grant No.2019M653849XB)the High-level Talents Introduction Plan of Yunnan Province-Young Talents Special Project。
文摘The formation of root system architecture(RSA)plays a crucial role in plant growth.OsDRO1 is known to have a function in controlling RSA in rice,however,the role of potato StDRO2,a homolog of rice OsDRO1,in root growth remains unclear.In this study,we obtained potato dro2 mutant lines by Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-Associated 9(CRISPR/Cas9)-mediated genome editing system.The mutant lines were generated from a splicing defect of the StDRO2 intron 1,which causes a nonsense mutation in StDRO2.Furthermore,the secondary structure of StDRO2 mRNA analyzed with RNAfold Web Server was altered in the dro2 mutant.Mutation of StDRO2 conveys potato adaptation through changing the RSA via alteration of auxin transport under drought stress.The potato dro2 lines showed higher plant height,longer root length,smaller root growth angle and increased tuber weight than the wild-type.The alteration of RSA was associated with a disturbance of IAA distribution in the dro2 mutant,and the levels of StPIN7 and StPIN10 detected by using real-time PCR were up-regulated in the roots of potato dro2 lines grown under drought stress.Moreover,the microRNAs(miRNAs)PmiREN024536 and PmiREN024486 targeted the StDRO2 gene,and auxin positively and negatively regulated the expression of StDRO2 and the miRNAs PmiREN024536 and PmiREN024486,respectively,in the potato roots.Our data shows that a regulatory network involving auxin,StDRO2,PmiREN024536 and PmiREN024486 can control RSA to convey potato fitness under drought stress.
基金funded by the Science and Technology Research Program of the Chongqing Municipal Education Commission(grant number KJQN202403002).
文摘The steel tube arch rib in a large-span concrete-filled steel tube arch bridge has a large span and diameter,which also leads to a larger weld seam scale.Large-scale welding seams will inevitably cause more obvious welding residual stress(WRS).For the purpose of studying the influence of WRS from large-scale welding seam on the mechanical properties of steel tube arch rib during arch rib splicing,test research and numerical simulation analysis on the WRS in arch rib splicing based on the Guangxi Pingnan Third Bridge,which is the world’s largest span concrete-filled steel tube arch bridge,were conducted in this paper,and the distribution pattern of WRS at the arch rib splicing joint was obtained.Subsequently,the WRS was introduced into the mechanical performance analysis of joints and structures to analyze its effects.The findings reveal that the distribution of WRS in the arch rib is greatly influenced by the rib plate,and the axial WRS in the heat-affected zone are primarily tensile,while the circumferential WRS are distributed in an alternating pattern of tensile and compressive stresses along the circumferential direction of the main tube.Under the influence of WRS,the ultimate bearing capacity of the joint is reduced by 29.4%,the initial axial stiffness is reduced by 4.32%,and the vertical deformation of the arch rib structure is increased by 4.7%.
基金Supported by the Jiangxi Provincial Health Technology Project(No.202130037 to CWL)was also sponsored by the Jiangxi Provincial Health Technology Project(No.202410009 to SHL)。
文摘Objectives:KH-type splicing regulatory protein(KHSRP)is an RNA-binding protein involved in several cellular processes,including nuclear splicing,mRNA localization,and cytoplasmic degradation.While KHSRP’s role has been studied in other cancers,its specific involvement in gastric cancer remains poorly understood.This study aims to explore KHSRP expression in gastric cancer and its potential effects on tumor progression and immune response.Methods:KHSRP expression in gastric cancer tissues and normal tissues was analyzed using data from The Cancer Genome Atlas(TCGA)database.The correlation between KHSRP expression,patient survival,and immune response was also assessed.Immunohistochemistry was performed to evaluate KHSRP expression in gastric cancer tissues.Gain-and loss-of-function experiments were conducted to assess KHSRP’s effects on gastric cancer cell proliferation,stemness,and migration.Furthermore,the impact of KHSRP silencing on tumor volume and immune cell infiltration was evaluated in a C3H/He mouse xenograft model.Results:KHSRP was found to be overexpressed in gastric cancer tissues compared to normal tissues,with a positive correlation to tumor stage and a negative correlation with patient prognosis.Functional assays revealed that KHSRP promotes gastric cancer cell proliferation,enhances cancer stem cell properties,and increases migratory capabilities in vitro.In vivo,KHSRP silencing led to a significant reduction in tumor volume and increased immune cell infiltration in the mouse xenograft model.Conclusions:KHSRP acts as an oncogene in gastric cancer by promoting tumorigenesis and suppressing anti-tumor immune responses.Its overexpression is associated with poor prognosis,making KHSRP a potential prognostic marker and therapeutic target in gastric cancer.
基金supported by the National Natural Science Foundation of China(32072126 and 32230075)the Shandong Provincial Natural Science Foundation(ZR2019MC005).
文摘Pentatricopeptide repeat(PPR)proteins are a large group of eukaryote-specific RNA-binding proteins that play pivotal roles in plant organelle gene expression.Here,we report the function of PPR21 in mitochondrial intron splicing and its role in maize kernel development.PPR21 is a typical P-type PPR protein targeted to mitochondria.The ppr21 mutants are arrested in embryogenesis and endosperm development,leading to embryo lethality.Null mutations of PPR21 reduce the splicing efficiency of nad2 intron 1,2,and 4 and impair the assembly and activity of mitochondrial complex I.Previous studies show that the P-type PPR protein EMP12 is required for the splicing of identical introns.However,our protein interaction analyses reveal that PPR21 does not interact with EMP12.Instead,both PPR21 and EMP12 interact with the small MutS-related(SMR)domain-containing PPR protein 1(PPR-SMR1)and the short P-type PPR protein 2(SPR2).PPR-SMR1 interacts with SPR2,and both proteins are required for the splicing of many introns in mitochondria,including nad2 intron 1,2,and 4.These results suggest that a PPR21-(PPR-SMR1/SPR2)-EMP12 complex is involved in the splicing of nad2 introns in maize mitochondria.
文摘In this study,we examine the problem of sliced inverse regression(SIR),a widely used method for sufficient dimension reduction(SDR).It was designed to find reduced-dimensional versions of multivariate predictors by replacing them with a minimally adequate collection of their linear combinations without loss of information.Recently,regularization methods have been proposed in SIR to incorporate a sparse structure of predictors for better interpretability.However,existing methods consider convex relaxation to bypass the sparsity constraint,which may not lead to the best subset,and particularly tends to include irrelevant variables when predictors are correlated.In this study,we approach sparse SIR as a nonconvex optimization problem and directly tackle the sparsity constraint by establishing the optimal conditions and iteratively solving them by means of the splicing technique.Without employing convex relaxation on the sparsity constraint and the orthogonal constraint,our algorithm exhibits superior empirical merits,as evidenced by extensive numerical studies.Computationally,our algorithm is much faster than the relaxed approach for the natural sparse SIR estimator.Statistically,our algorithm surpasses existing methods in terms of accuracy for central subspace estimation and best subset selection and sustains high performance even with correlated predictors.
基金Supported by National Natural Science Foundation of China,No.82160762Guangxi Zhuang Autonomous Region Administration of Traditional Chinese Medicine Scientific Research Project,No.GXZYA20230267+2 种基金China Undergraduate Innovation and Entrepreneurship Training Program,No.S202410598060XChina Undergraduate Innovation and Entrepreneurship Training Program,No.X202410598360Future Academic Star of Guangxi Medical University,No.WLXSZX24074.
文摘BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on the evaluation of the potential role of PSMD6 in hepatocellular carcinoma(HCC).AIM To comprehensively evaluate the overexpression pattern and clinical significance of PSMD6 in HCC tissues.METHODS This study integrated PSMD6 mRNA expression profiles from 4672 HCC and 3667 non-HCC tissues,along with immunohistochemical scores from 383 HCC and adjacent tissues,to assess PSMD6 overexpression in HCC.Clustered regularly interspaced short palindromic repeats knockout technology evaluated PSMD6’s essential role in HCC cell growth.Functional enrichment analysis explored the molecular mechanism of PSMD6 abnormalities in HCC.Drug sensitivity analysis and molecular docking analysed the effect of abnormal expression of PSMD6 on the drug sensitivity of HCC cells.RESULTS The results of 41 external and two internal datasets showed that PSMD6 mRNA(SMD=0.26,95%CI:0.09-0.42,P<0.05)and protein(SMD=2.85,95%CI:1.19-4.50,P<0.05)were significantly overexpressed in HCC tissues.The integrated analysis results showed that PSMD6 had a significant overexpression pattern in HCC tissues(SMD=0.40,95%CI:0.15-0.66,P<0.05).PSMD6 knockout inhibited HCC cell growth(chronos scores<-1).Functional enrichment implicated ribosome biogenesis and RNA splicing.Significant enrichment of signalling pathways such as RNA degradation,ribosomes,and chemical carcinogenesis—reactive oxygen species.Drug sensitivity analysis and a molecular docking model showed that high expression of PSMD6 was associated with the tolerance of HCC cells to drugs such as ML323,sepantronium bromide,and GDC0810.Overexpressed PSMD6 effectively distinguished HCC tissues(AUC=0.75,95%CI:0.71-0.79).CONCLUSION This study was the first to discover that PSMD6 was overexpressed in HCC tissues.PSMD6 is essential for the growth of HCC cells and may be involved in ribosome biogenesis and RNA splicing.
基金support provided by the National Natural Science Foundation of China(No.52002324)the Hong Kong Scholars Program(Nos.XJ2021073 and PolyU YZ4V)the Research Grants Council of Hong Kong GRF(No.15303123).
文摘Stimulus-responsive actuators are novel functional devices capable of sensing external stimuli and ex-hibiting specific deformation responses.MXene,owing to its unique 2D structure and efficient energy conversion efficiency,has bridged the gap in traditional devices and shown great potential for multiple stimulus-responsive actuators.However,the drawbacks of pure MXene films,including susceptibility to oxidation and vulnerability to shear stress,hinder their applications.Through composite modification and structural design strategies,a three-layer structured MXene-carbon nanotubes hybrid film(tHCM)is fabri-cated,exhibiting a tensile strength and fracture strain of 153.8 MPa and 4.65%,respectively,representing improvements of 598.4%and 226.8%compared to the initial film.Meanwhile,the film maintains excel-lent stability demonstrating the enhancing effects of hydrogen bonds and densely packed structure.The hybrid films demonstrate unique and facile welding features due to splicing properties,enabling the for-mation of complex configurations.In terms of electro-/photo-thermal conversion performance,the hybrid film can reach a reasonably high temperature of 250℃at low voltage(2.5 V)and 110.6℃under 150 mW cm^(-2) infrared light.Leveraging the thermal expansion mismatch between tHCM and thermoplastic films,an integrated,flexible,and weldable actuator with unique electro/photo-response is developed,and vari-ous biomimetic driving applications,particularly,the light-mediated hierarchical transmission and precise motion along predetermined trajectory are realized.This work not only provides an effective strategy for modifying MXene composite films but also advances the design of novel actuators,offering broad appli-cation prospects in fields such as stimulus-responsive actuated robots and cargo transportation.
基金funded by the National Natural Science Foundation of China(32202393)the Natural Science Foundation of Shandong Province,China(ZR2021QC190)+1 种基金the Science and Technology Benefiting the People Demonstration Project of Qingdao,China(24-1-8-xdny-10-nsh)the Qingdao Agricultural University High-level Talent Fund,China(663/1120101)。
文摘Caspases,which play key roles in cell apoptosis,undergo alternative splicing to form different splicing variants that can regulate the apoptotic process.Lepidopteran insect caspases undergo alternative splicing,although the functions of their splicing variants are still unclear.The Spodoptera exigua caspase-5(SeCaspase-5)gene was cloned and found to produce four different splicing variants with different gene sequences and protein functional domains,which were named SeCaspase-5a,SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Overexpression of these variants in S.exigua cells(Se-3)showed that SeCaspase-5a had a proapoptotic function,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not.Semi-qPCR analysis revealed that the expression of the SeCaspase-5 variants significantly differed during Autographa californica multiple nucleopolyhedrovirus(AcMNPV)infection.Furthermore,the SeCaspase-5 variants were constructed into the AcMNPV bacmid and transfected into Se-3 cells,which revealed that SeCaspase-5a promoted cell apoptosis and reduced virus production,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not promote cell apoptosis but instead increased virus production.Moreover,an analysis of the interactions between the SeCaspase-5 variants revealed that SeCaspase-5a directly interacted with SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Coexpression of these variants in Se-3 cells also revealed that SeCaspase-5b,SeCaspase-5c and SeCaspase-5d inhibited the proapoptotic function of SeCaspase-5a,resulting in a reduction in the percentage of apoptotic cells by about 20%.These results indicate that SeCaspase-5 undergoes alternative splicing and is involved in regulating the apoptosis induced by baculovirus infection.These findings increase our understanding of the functions of lepidopteran insect caspases and provide new insights into the mechanism of host-cell apoptosis induced by baculoviruses.
基金supported by the National Nature Science Foundation of China(81902179)the Postdoctoral Science Foundation of China(2020T130308)+3 种基金the Key Medical Discipline of Jiangsu Province(JSDW202223)the Natural Science Foundation of Jiangsu Province(BK20221241)the Science and Technology Project of Suzhou(SKJY2021094)the Gusu Talent Program(GSWS2022046)。
文摘Survival of motor neuron(SMN)protein encoded by SMN1 gene,is the essential and ubiquitously expressed protein in all tissues.Prior studies demonstrated that SMN deficiency impaired bone development,but the underlying mechanism of abnormal endochondral ossification remains obscure.Here,we showed SMN is involved in hypertrophic chondrocytes differentiation through regulating RNA splicing and protein degradation via analyzing single cell RNA-sequencing data of hypertrophic chondrocytes.Of note,SMN loss induced dwarfism and delayed endochondral ossification in Smn1 depletion-severe spinal muscular atrophy(SMA)mouse model and Smn1 chondrocyte conditional knockdown mouse.Histological analysis revealed that SMN deficiency expanded the zone of hypertrophic chondrocytes in the growth plates,but delayed turnover from hypertrophic to ossification zone.Widespread changes in endochondral ossification related gene expression and alternative splicing profiles were identified via RNA sequencing of growth plate cartilages from SMA mice on postnatal day 4.Importantly,Mass spectrometry-based proteomics analysis elucidated Y-box-binding protein 1(YBX1)as a vital SMN-binding factor,was decreased in SMA mice.YBX1 knockdown reproduced the aberrant gene expression and splicing changes observed in SMA growth plate cartilages.Comparing the binding proteins of SMN and YBX1 revealed TNF receptor-associated factor 6(TRAF6),which promoted ubiquitination degradation of YBX1.By conditionally deleting Smn1 in chondrocytes of WT mice and overexpressing Smn1 in chondrocytes of SMA mice,we proved that SMN expression in chondrocytes is critical for hypertrophic chondrocyte-mediated endochondral ossification.Collectively,these results demonstrate that SMN deficiency contributes to rapid systemic bone dysplasia syndrome by promoting TRAF6-induced ubiquitination degradation of YBX1 in growth plate cartilages of SMA mice.
基金supported by the National Natural Science Foundation of China(32130098)。
文摘Male infertility constitutes a major global public health concern,with the underlying etiology remaining unidentified in nearly half of the diagnosed cases.Protein kinase CK1α(CK1α)functions as a pivotal regulator of cell cycle progression,pre-mRNA processing,and spliceosome-associated pathways through interactions with specific splicing factors.Comprehensive analyses revealed CK1αexpression in both germ cells and somatic cells of mouse testes,implicating its involvement in spermatogenic regulation.However,the physiological roles and mechanistic basis of CK1αfunction in Sertoli cells remain unclear.In this study,CK1αwas highly expressed in Sertoli cells,and conditional knockout of CK1αin murine Sertoli cells induced profound testicular atrophy and complete infertility.This phenotype was characterized by rapid depletion of Sertoli cells and spermatogenic dysfunction.Subsequent analyses demonstrated that CK1αregulated the fate determination of fetal and neonatal Sertoli cells in mice.At the molecular level,CK1αpromoted Sertoli cell survival through interaction with the splicing factor ZRSR1 to modulate apoptosis.Collectively,these findings establish CK1αas a key regulator of alternative splicing and male reproduction,providing critical insights into the molecular mechanisms underlying testicular development and reproductive function.
基金supported in part by the Natural Science Foundation of Shandong Province(no.ZR2022QH373,ZR2022QH292 and ZR2023MH2474).
文摘ObjectivesThe PTPRQ gene is essential for preserving the structure and function of stereocilia in inner ear.However,research on splicing mutations within this gene is limited.This study aims to investigate novel splicing mutations in PTPRQ,clarify their molecular mechanisms,and provide new insights into the genetic factors associated with hearing loss,ultimately enhancing diagnostic accuracy.MethodClinical data and peripheral blood samples were obtained from members of a family with congenital hearing loss.Variants were identified through high-throughput sequencing and confirmed by Sanger sequencing to ensure genealogical co-segregation.The splicing effects of PTPRQ variants were evaluated using bioinformatics tools and minigene assays.ResultsWe used whole exome sequencing to identify novel double compound heterozygous splice-altering variants(c.5426+1 G>A and c.6603-3 T>G)in the PTPRQ gene with DFNB84A.We molecularly characterized these variants,and they were found to co-segregate with the disease within the family.Minigene assays and Sanger sequencing confirmed that the c.6603-3 T>G variant caused exon 43 skipping,resulting in a frameshift mutation(p.Ser2201ArgfsTer112).Further bioinformatic analysis supported these findings.ConclusionsThis study identifies a novel compound heterozygous splicing variant in the PTPRQ gene in a Chinese family with DFNB84A,expanding the known spectrum of PTPRQ mutations.These findings enhance the understanding of PTPRQ-related hearing loss and may aid in early diagnosis,prevention,and therapeutic strategies.
基金supported by the Science and Technology Program of Hebei Province, China (236Z2903G)the Innovative Research Group Project of Hebei Natural Science Foundation, China (C2024204246)+1 种基金the Hebei International Joint Research Center of Vegetable Functional Genomicsthe International Joint R&D Center of Hebei Province in Modern Agricultural Biotechnology for supporting this work。
文摘The plant circadian clock temporally drives gene expression throughout the day and coordinates various physiological processes with diurnal environmental changes. It is essential for conferring plant fitness and competitive advantages to survive and thrive under natural conditions through the circadian control of gene transcription. Chinese cabbage(Brassica rapa ssp. pekinensis) is an economically important vegetable crop worldwide, although there is little information concerning its circadian clock system. Here we found that gene expression patterns are affected bycircadian oscillators at both the transcriptional and post-transcriptional levels in Chinese cabbage. Time-course RNA-seq analyses were conducted on two short-period lines(SPcc-1 and SPcc-2) and two long-period lines(LPcc-1 and LPcc-2) under constant light. The results showed that 32.7–50.5% of the genes were regulated bythe circadian oscillator and the expression peaks of cycling genes appeared earlier in short-period lines than long-period lines. In addition, approximately 250 splicing events exhibited circadian regulation, with intron retention(IR) accounting for a large proportion. Rhythmically spliced genes included the clock genes LATE ELONGATEDHYPOCOTYL(BrLHY), REVEILLE 2(BrRVE2) and EARLY FLOWERING 3(BrELF3). We also found that thecircadian oscillator could notably influence the diurnal expression patterns of genes that are associated with glucose metabolism via photosynthesis, the Calvin cycle and the tricarboxylic acid(TCA) cycle at both the transcriptional andpost-transcriptional levels. The collective results of this study demonstrate that circadian-regulated physiological processes contribute to Chinese cabbage growth and development.
基金supported by the National Natural Science Foundation of China(Nos.81972771,82173062)the Key Areas Project of Education Department of Guangdong Province(No.2021ZDZX2017)+3 种基金the Tertiary Education Scientific Research Project of Guangzhou Municipal Education Bureau(No.202235387)the Guangzhou Science and Technology Project of Guangzhou Municipal Science and Technology Bureau(No.2023A03J0428)the Natural Science Foundation of Guangdong Province,China(No.2024A1515013082)the Guangdong Basic and Applied Basic Research 21 Foundation(No.2021A1515010403).
文摘Background:Alterations in splicing factors contribute to aberrant alternative splicing(AS),which subsequently promotes tumor progression.The splicing factor polypyrimidine tract binding protein 1(PTBP1)has been shown to facilitate cancer progression by modulating oncogenic variants.However,its specific role and underlying mechanisms in hepatocellular carcinoma(HCC)remain to be elucidated.Methods:PTBP1 expression was evaluated in HCC tissues and cell lines.Subsequently,cells were transfected with vectors designed for PTBP1 overexpression or downregulation.The biological function of PTBP1 was assessed in vitro and in vivo using MTS assays,colony formation assays,transwell assays,xenograft formation,tail vein injection,and orthotopic models.Transcriptome analysis was conducted to elucidate the underlying molecular mechanisms.Results:Our findings demonstrated that PTBP1 exhibited elevated expression in HCC cell lines and tissues.Furthermore,its expression positively correlated with overall and disease-free survival rates,as well as tumor grade and stage.PTBP1 knockdown reduced HCC cell proliferation,migration,and invasion in vitro and suppressed hepatocarcinoma xenograft growth and infiltration in vivo.RNA sequencing(RNA-Seq)analysis identified the AS events associated with PTBP1.PTBP1 functionally enhanced cell proliferation,invasion,and migration by modulating the AS of the microtubule-associated protein tau(MAPT)gene and promoting oncogene expression.Notably,the dysregulation of MAPT splicing coincided with increased PTBP1 expression in HCC.Conclusions:PTBP1-guided AS of the MAPT gene enhances tumorigenicity in HCC through activation of the MAPK/ERK pathways.
基金supported by the National Key Research and Development Program of China(2021YFF1000301)the National Natural Science Foundation of China(31771805)。
文摘Heat stress is a major threat to maize(Zea mays L.)production worldwide.Heat shock transcription factors(HSFs)play vital roles in plant responses to heat stress.However,the molecular and genetic mechanisms underlying HSF-meditated thermotolerance in maize remain largely unexplored.In this study,we demonstrate that the alternative splicing of ZmHsf23 modulates heat stress tolerance in maize.Hsf23 produced two functional transcripts,Hsf23L and Hsf23S,which differ by the presence of a cryptic mini-exon in Hsf23L that is spliced out in Hsf23S.Both transcripts were strongly induced by heat stress.Mutants lacking Hsf23L alone(hsf23l)or both Hsf23L and Hsf23S(hsf23l23s)exhibited increased susceptibility to heat stress,whereas overexpression of Hsf23S enhanced heat stress tolerance in maize.Subsequently,we found that Hsf23S positively regulates heat stress tolerance by directly activating the transcription of three sHSP genes(Hsp16.9,Hsp17.2,and Hsp18a)and TIL1 gene.In addition,Hsf23L physically interacted with Hsf23S and enhanced the transcriptional activation of Hsf23S on the sHSPs and TIL1 promoters.Notably,genetic analysis suggested that co-overexpression of Hsf23L and Hsf23S further improves heat tolerance of the transgenic plants.Taken together,these results reveal two splicing variants of ZmHsf23 cooperatively regulate maize heat tolerance,thus highlighting potential value of ZmHsf23 in breeding heat-tolerant maize varieties.
基金supported by the Major Program of National Agricultural Science and Technology of China(NK20220607)the Sichuan Science and Technology Program,China(2023YFH0041)。
文摘Starch biosynthesis is a complex process that relies on the coordinated action of multiple enzymes.Resistant starch is not digested in the small intestine,thus preventing a rapid rise in the glycemic index.Starch synthase 2a(SS2a)is a key enzyme in amylopectin biosynthesis that has significant effects on starch structure and properties.In this study,we identified an ss2a null mutant(M3-1413)with a single base mutation from an ethyl methane sulfonate(EMS)-mutagenized population of barley.The mutation was located at the 3'end of the first intron of the RNA splicing receptor(AG)site,and resulted in abnormal RNA splicing and two abnormal transcripts of ss2a,which caused the inactivation of the SS2a gene.The starch structure and properties were significantly altered in the mutant,with M3-1413 containing lower total starch and higher amylose and resistant starch levels.This study sheds light on the effect of barley ss2a null mutations on starch properties and will help to guide new applications of barley starch in the development of nutritious food products.
基金supported by the Hainan Province Science and Technology Talent Innovation Project(KJRC2023A02)Project of Sanya Yazhouwan Science and Technology City Management Foundation(SKJC-KJ-2019KY01)Sanya Science and Technology Special Fund(2022KJCX91)。
文摘The silver-lipped pearl oyster(Pinctada maxima)is the largest and most commercially valuable pearl-producing oyster,renowned for its ability to generate large,lustrous pearls.This species is a sequential hermaphrodite,with pearl production displaying notable sexual dimorphism.Consequently,understanding the molecular mechanisms governing sex determination and differentiation is crucial for advancing breeding strategies in the pearl oyster industry.To elucidate these mechanisms,this study conducted integrative transcriptomic analyses of P.maxima gonadal tissues using isoform sequencing(Isoseq)and RNA sequencing(RNA-seq).Comparative analysis of ovarian and testicular tissues identified 2768 differentially expressed genes(DEGs).Gene coexpression network analysis delineated four key modules,including three sex-specific modules and one shared module.Key genes implicated in sex determination and maintenance were identified,including FOXL2,NANOS1,andβ-catenin,important for ovarian maintenance,and DMRT,SOX30,FEM1,and FOXJ1,crucial for testicular maintenance.These genes,widely studied in other taxa,were confirmed as hub genes in the sex-related modules of P.maxima.Interestingly,genes within the shared module were significantly enriched in the spliceosome pathway.Alternative splicing analysis highlighted its extensive role in gonadal tissues,with more pronounced activity observed in the testis compared to the ovary.Nearly half(47.83%,375)of the identified genes undergoing differential alternative splicing(DASGs)also exhibited differential transcript usage(DTUGs),while only 17%of DTUGs overlapped with DEGs.Genes associated with sex differentiation,such as DMRT,β-catenin,and U2AF2,displayed sex-specific and/or sex-biased isoforms.These findings offer novel insights into the molecular basis of sex differentiation in P.maxima,which could inform the development of targeted breeding strategies aimed at sex control,thereby enhancing pearl quality and yield in aquaculture.This study offers a robust molecular foundation for advancing breeding programs and optimizing production in the pearl oyster industry.
基金funded by Guizhou Province Science and Technology Plan Project Qiankehe Foundation-ZK[2023]General 360,362Science and Technology Fund project of Guizhou Provincial Health Commission(gzwkj-2022-09,gzwkj-2023-035)+1 种基金National Natural Science Foundation Cultivation Project of Guizhou Medical University(21NSFCP14,gyfynsfc-2022-25)The PhD Scientific Research Launch Fund Project of the Affiliated Hospital of Guizhou Medical University(gyfybsky-2022-02).
文摘Background:Glioblastoma(GBM)is one of the most malignant types of central nervous system tumors.Oxygen deprivation in the tumor microenvironment is thought to be an important factor in promoting GBM progression.However,the mechanisms of hypoxia-promoted tumor progression remain elusive.Methods:Alternative splicing of diacylglycerol kinase gamma(DGKG)-Δexon13 was amplified and verified by PCR-Sanger sequencing.The functions of DGKG and DGKG-Δexon13 were analyzed by Cell counting kit-8(CCK-8),Transwell,Matrigeltranswell experiments,and in vivo orthotropic GBM animal models.Transcriptome analyses were done to find out the regulated genes.Results:In this study,we found that a new transcript DGKG-Δexon13 was generated in GBM under hypoxia via alternative splicing.Moreover,the results of CCK-8,Transwell,and Matrigel-transwell experiments showed that the proliferation,migration,and invasion abilities of U87-MG and T98G were decreased after DGKG knockdown.Compared to wild-type DGKG,DGKG-Δexon13 overexpression significantly promoted cellular proliferation,migration,and invasion abilities in GBM.Furthermore,in vivo,orthotropic GBM animal models analysis showed that the tumor volumes were much smaller in the DGKG knockdown group.However,the tumor sizes in the DGKG and DGKG-Δexon13 rescue groups were restored,especially in the DGKG-Δexon13 group.Transcriptome analysis revealed that MORC1,KLHDC7B,ATP1A2,INHBE,TMEM119,and FGD3 were altered significantly when DGKG was knocked down.IL-16,CCN2,and EFNB3 were specifically regulated by DGKG-Δexon13.Conclusions:Our study found that hypoxia-induced alternative splicing transcript DGKG-Δexon13 promotes GBM proliferation and infiltration,which might provide a new potential target for the clinical treatment and diagnosis of GBM.
基金supported by the Jiangsu Provincial Key Research and Development Program(BE2022072)the National Natural Science Foundation of China(12141304)the Natural Science Foundation of Jiangsu Province(BK20231134).
文摘To address the installation challenges of a 2-m ring Gregorian telescope system,and similar optical systems with a small width-to-radius ratio,we propose a detection method combining local interferometry with a comparison model.This method enhances the precision of system calibration by establishing a dataset that delineates the relationship between secondary mirror misalignment and wavefront aberration,subsequently inferring the misalignment from interferometric detection results during the calibration process.For the 2-m ring telescope,we develop a detection model using five local sub-apertures,enabling a root-mean-square detection accuracy of 0:0225λ(λ=632:8 nm)for full-aperture wavefront aberration.The calibration results for the 2-m Ring Solar Telescope system indicate that the root-mean-square value of sub-aperture wavefront aberration reaches 0.104λ,and the root-mean-square value of spliced full-aperture measurement yields reaches 0.112λ.This method offers a novel approach for calibrating small width-toradius ratio telescope systems and can be applied to the calibration of other irregular-aperture optical systems.
