Objective:Polycystic ovary syndrome(PCOS)is a common endocrine disorder that affects women’s health.This study aims to investigate gene and transcription factor(TF)expression differences between PCOS patients and hea...Objective:Polycystic ovary syndrome(PCOS)is a common endocrine disorder that affects women’s health.This study aims to investigate gene and transcription factor(TF)expression differences between PCOS patients and healthy individuals using bioinformatics approaches,and to verify the function of key transcription factors,with the goal of providing new insights into the pathogenesis of PCOS.Methods:Differentially expressed genes(DEGs)and differentially expressed transcription factors(DETFs)between PCOS patients and controls were identified from the RNA sequencing dataset GSE168404 using bioinformatics methods.Functional enrichment analysis was performed using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases.The expression and function of core transcription factors were further validated in ovarian tissues of PCOS model mice and control mice using Western blotting and reverse transcription quantitative polymerase chain reaction(RTqPCR).Results:A total of 332 DEGs were identified between PCOS patients and controls,including 259 upregulated and 73 downregulated genes in the PCOS group.19 DETFs were further screened,of which 16 were upregulated and 3 were downregulated in PCOS.The upregulated DETFs(including TFCP2L1,DACH1,ESR2,AFF3,SMAD9,ZNF331,HOPX,ATOH8,HIF3α,DPF3,HOXC4,HES1,ID1,JDP2,SOX4,and ID3)were primarily associated with lipid metabolism,development,and cell adhesion.Protein and mRNA expression analysis in PCOS model mice revealed significantly decreased levels of hypoxia-inducible factor(HIF)1αand HIF2α,and significantly increased expression of HIF3αcompared to control mice(all P<0.001).Conclusion:Significant differences in gene and TF expression exist between PCOS patients and healthy individuals.HIF-3αmay play a crucial role in PCOS and could serve as a novel biomarker for diagnosis and a potential therapeutic target.展开更多
Drought is one of the most important environmental constraints limiting plant growth, development and crop yield. Many drought-inducible genes have been identified by molecular and genomic analyses in Arabidopsis, ric...Drought is one of the most important environmental constraints limiting plant growth, development and crop yield. Many drought-inducible genes have been identified by molecular and genomic analyses in Arabidopsis, rice and other crops. To better understand reaction mechanism of plant to drought tolerance, we mainly focused on introducing the research of transcription factors （TFs） in signal transduction and regulatory network of gene expression conferring drought. A TF could bind multiple target genes to increase one or more kinds of stress tolerance. Sometimes, several TFs might act together with a target gene. So drought-tolerance genes or TFs might respond to high-salinity, cold or other stresses. The crosstalk of multiple stresses signal pathways is a crucial aspect of understanding stress signaling.展开更多
Abiotic stress including drought,lowtemperature,ABA and high salt is major factoraffecting the plant growth.Isolation andfunctional study of abiotic stress-related genewill be helpful to elucidate the signaltransducti...Abiotic stress including drought,lowtemperature,ABA and high salt is major factoraffecting the plant growth.Isolation andfunctional study of abiotic stress-related genewill be helpful to elucidate the signaltransduction mechanism of the target gene underabiotic stress during the growth of plant.Byusing gene-transfer technique,the target gene isincorporated into the plant to improve theadaptation of plant to abiotic stress.展开更多
Climate deterioration,water shortages,and abiotic stress are the main threats worldwide that seriously affect cotton growth,yield,and fiber quality.Therefore,research on improving cotton yield and tolerance to biotic ...Climate deterioration,water shortages,and abiotic stress are the main threats worldwide that seriously affect cotton growth,yield,and fiber quality.Therefore,research on improving cotton yield and tolerance to biotic and abiotic stresses is of great importance.The NAC proteins are crucial and plant-specific transcription factors(TFs)that are involved in cotton growth,development,and stress responses.The comprehensive utilization of cotton NAC TFs in the improvement of cotton varieties through novel biotechnological methods is feasible.Based on cotton genomic data,genome-wide identification and analyses have revealed potential functions of cotton NAC genes.Here,we comprehensively summarize the recent progress in understanding cotton NAC TFs roles in regulating responses to drought,salt,and Verticillium wilt-related stresses,as well as leaf senescence and the development of fibers,xylem,and glands.The detailed regulatory network of NAC proteins in cotton is also elucidated.Cotton NAC TFs directly bind to the promoters of genes associated with ABA biosynthesis and secondary cell-wall formation,participate in several biological processes by interacting with related proteins,and regulate the expression of downstream genes.Studies have shown that the overexpression of NAC TF genes in cotton and other model plants improve their drought or salt tolerance.This review elucidates the latest findings on the functions and regulation of cotton NAC proteins,broadens our understanding of cotton NAC TFs,and lays a fundamental foundation for further molecular breeding research in cotton.展开更多
AIM:Normalizing the results of real-time quantitative reverse transcription polymerase chain reaction(RT-q PCR)is essential for the accuracy of analysis.Commonly used approaches include input nucleic acid standardizat...AIM:Normalizing the results of real-time quantitative reverse transcription polymerase chain reaction(RT-q PCR)is essential for the accuracy of analysis.Commonly used approaches include input nucleic acid standardization(ΔCt method),normalization against a single internal reference gene(ΔΔCt method),and geometric averaging of multiple reference gene abundance using statistical software.We evaluated these approaches in the liver of db/db mice,a typical model of fatty liver disease.METHODS:Seven reference genes,β-actin(ACTB),eukaryotic initiation factor(e IF)5,glyceraldehyde-3-phosphate dehydrogenase(GAPDH),hydroxymethylbilane synthase(HMBS),hypoxanthineguanine phosphoribosyltransferase(HPRT)1,polymerase(RNA)II(DNA directed)polypeptide A(Polr2A)and ribosomal protein P〈0(RPLP〈0),were evaluated using software of ge Norm and Norm Finder.Hepatic lipogenesis genes,such as thyroid hormone-responsive protein(Thrsp),stearoyl-Co A desaturase(SCD)1,sterol regulatory element-binding protein(SREBP)1c and fatty acid synthase(FAS),were used as target genes of interest.RESULTS:The expression levels of all target genes and GAPDH were significantly elevated(P〈0.05)in db/db mouse livers by theΔCt method.ACTB and HMBS were the most stable genes calculated by the software of ge Norm.Norm Finder analysis indicated that ACTB was the most stable gene,and the best combination of 2 genes was GAPDH and RPLP〈0.Normalization against a single internal reference gene of ACTB or RPLP〈0,the geometric mean of ACTB and HMBS,or GAPDH and RPLP〈0 showed similar results that the expression levels of Thrsp,SCD1 and FAS,but not SREBP1 c increased(P〈0.05)in the liver of db/db mice.CONCLUSION:TheΔCt approach ensures a meaningful and biologically significant appraisal of gene expression.Use of the software like ge Norm or Norm Finder should be integrated withΔCt method.展开更多
The myeloblastosis oncogenes(MYB)are important transcription factors that facilitate induction of variously develop-mental and stress responsive genes.They are hence,emerging as key players in improving stress toleran...The myeloblastosis oncogenes(MYB)are important transcription factors that facilitate induction of variously develop-mental and stress responsive genes.They are hence,emerging as key players in improving stress tolerance of plants in response to several abiotic stresses.It was predicted that DfMYB2 contained an open reading frame(ORF)of 1621 bp coding 377 amino acid residues with molecular weight of 41.5 ku.It had 50 potential phosphorylation sites,44 potential N-glycosylation sites and one trans-membrane domain.In neighbor-joining(NJ)phylogenetic tree,DfMYB2 was close to the branch of angiosperms.The subcellular localization of DfMYB2 was in nucleus.The expressions of DfMYB2 increased gradually in the filament,prothalli and sporophyte.The results showed that DfMYB2 played a more and more important role in its growth and development.And the expression levels in rhizomes were significantly higher than those in roots and leaves.After abscisic acid(ABA)and PEG600 treatment,DfMYB2 showed a downward trend,followed by an upward trend.The expression of DfMYB2 was inhibited in a short time under drought stress,the strong induction of DfMYB2 expression by ABA indicated that it was possibly involved in abiotic stress responses in an ABA-dependent manner.展开更多
Background:INDETERMINATE DOMAIN(IDD)transcription factors form one of the largest and most conserved gene families in plant kingdom and play important roles in various processes of plant growth and development,such as...Background:INDETERMINATE DOMAIN(IDD)transcription factors form one of the largest and most conserved gene families in plant kingdom and play important roles in various processes of plant growth and development,such as flower induction in term of flowering control.Till date,systematic and functional analysis of IDD genes remained infancy in cotton.Results:In this study,we identified total of 162 IDD genes from eight different plant species including 65 IDD genes in Gossypium hirsutum.Phylogenetic analysis divided IDDs genes into seven well distinct groups.The gene structures and conserved motifs of GhIDD genes depicted highly conserved exon-intron and protein motif distribution patterns.Gene duplication analysis revealed that among 142 orthologous gene pairs,54 pairs have been derived by segmental duplication events and four pairs by tandem duplication events.Further,Ka/Ks values of most of orthologous/paralogous gene pairs were less than one suggested the purifying selection pressure during evolution.Spatiotemporal expression pattern by qRT-PCR revealed that most of the investigated GhIDD genes showed higher transcript levels in ovule of seven days post anthesis,and upregulated response under the treatments of multiple abiotic stresses.Conclusions:Evolutionary analysis revealed that IDD gene family was highly conserved in plant during the rapid phase of evolution.Whole genome duplication,segmental as well as tandem duplication significantly contributed to the expansion of IDD gene family in upland cotton.Some distinct genes evolved into special subfamily and indicated potential role in the allotetraploidy Gossypium hisutum evolution and development High transcript levels of GhIDD genes in ovules illustrated their potential roles in seed and fiber development Further,upregulated responses of GhIDD genes under the treatments of various abiotic stresses suggested them as important genetic regulators to improve stress resistance in cotton breeding.展开更多
Background:Pectin is a key substance involved in cell wall development,and the galacturonosyltransferases(GAUTs)gene family is a critical participant in the pectin synthesis pathway.Systematic and comprehensive resear...Background:Pectin is a key substance involved in cell wall development,and the galacturonosyltransferases(GAUTs)gene family is a critical participant in the pectin synthesis pathway.Systematic and comprehensive research on GAUTs has not been performed in cotton.Analysis of the evolution and expression patterns of the GAUT gene family in different cotton species is needed to in crease kno wledge of the functi on of pectin in cotto n fiber development.Results:In this study,we have identified 131 GAUT genes in the genomes of four Gossypium species(G.raimondii,G barbadense,G.hirsutum,and G.arboreum),and classified them as GAUT-A,GAUT-B and GAUT-C,which coding probable galacturonosyltransferases.Among them,the GAUT genes encode proteins GAUT1 to GAUT15.All GAUT proteins except for GAUT7 contai n a con served glycosyl transferase family 8 domain(H-DN-A-SW-S-V-H-T-F).The conserved sequence of GAUT7 is PLN(phospholamban)02769 domain.According to c/s-elemet analysis,GAUT genes transcript levels may be regulated by horm ones such as JA,GA,SA,ABA,Me-JA,and IA A.The evoluti on and transcription patterns of the GAUT gene family in different cotton species and the transcript levels in upland cotton lines with different fiber st「ength were analyzed.Peak transcript level of GhGAUT genes have been observed before 15 DPA.In the six materials with high fiber strength,the transcription of GhGAUT genes were concentrated from 10 to 15 DPA;while the highest transcript levels in low fiber st「ength materials were detected between 5 and 10 DPA.These results lays the foundation for future research on gene function during cotton fiber development.Conclusions:The GAUT gene family may affect cotton fiber development,including fiber elongation and fiber thickening.In the low strength fiber lines,GAUTs mainly participate in fiber elongation,whereas their major effect on cotton with high strength fiber is related to both elongation and thickening.展开更多
Background:Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries globally.The effects of drought and salt stress pose a challenge to strong fiber and large-scale pr...Background:Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries globally.The effects of drought and salt stress pose a challenge to strong fiber and large-scale production due to the ever-changing climatic conditions.However,plants have evolved a number of survival strategies,among them is the induction of various stress-responsive genes such as the ribosomal protein large(RPL)gene.The RPL gene families encode critical proteins,which alleviate the effects of drought and salt stress in plants.In this study,comprehensive and functional analysis of the cotton RPL genes was carried out under drought and salt stresses.Results:Based on the genome-wide evaluation,26,8,and 5 proteins containing the RPL14B domain were identified in Gossypium hirsutum,G.raimondii,and G.arboreum,respectively.Furthermore,through bioinformatics analysis,key cis-regulatory elements related to RPL14B genes were discovered.The Myb binding sites(MBS),abscisic acid-responsive element(ABRE),CAAT-box,TATA box,TGACG-motif,and CGTCA-motif responsive to methyl jasmonate,as well as the TCA-motif responsive to salicylic acid,were identified.Expression analysis revealed a key gene,Gh_D01G0234(RPL14B),with significantly higher induction levels was further evaluated through a reverse genetic approach.The knockdown of Gh_D01G0234(RPL14B)significantly affected the performance of cotton seedlings under drought/salt stress conditions,as evidenced by a substantial reduction in various morphological and physiological traits.Moreover,the level of the antioxidant enzyme was significantly reduced in VIGS-plants,while oxidant enzyme levels increased significantly,as demonstrated by the higher malondialdehyde concentration level.Conclusion:The results revealed the potential role of the RPL14B gene in promoting the induction of antioxidant enzymes,which are key in oxidizing the various oxidants.The key pathways need to be investigated and even as we exploit these genes in the developing of more stress-resilient cotton germplasms.展开更多
Aim Inducible nitric oxide synthase （iNOS） makes a great contribution to host defense and inflamma-tion. In many settings, lipopolysaccharide （LPS） induces iNOS expression through activation of the inhibitor of K...Aim Inducible nitric oxide synthase （iNOS） makes a great contribution to host defense and inflamma-tion. In many settings, lipopolysaccharide （LPS） induces iNOS expression through activation of the inhibitor of KB- α （IKB-α） -nuclear factor-KB （NF-KB） cascade, whereas interferon-γ （IFN-γ） acts through Janus kinase （ JAK）- signal transducer and activator of transcription 1 （ STAT1 ） signals. Heat shock factor 1 （ HSF1 ）, a major regulator of heat shock protein transcription, has been shown to regulate the production of pro-inflammatory cytokines such as tumor necrosis factor-α（TNF-α） and interleukin-6 （IL-6）. But it remains obscure whether and how HSF1 affects iNOS induction. Methods Western blot was used to measure the protein expression. The mRNA level was meas- ured by real time-PCR. Silence of HSF1 was achieved by small interfering RNA. Nitric oxide （NO） content and NF-KB binding activity were assayed by commercial kits. Chromatin immunoprecipitation （CHIP） was used to measure the binding activity of NF-KB and STAT1 to iNOS promoters. Results HSF1 inhibition or knockdown pre- vented the LPS- and/or IFN-γ-stimulated iNOS protein expression in cultured microglia. HSF1 inhibition blocked iNOS mRNA transcription. These inhibitory effects of HSF1 inhibition on iNOS expression were confirmed in brain tissues from endotoxemic mice. Further analysis showed that HSF1 inhibition had no effect on IKB-α degradation and NF-KB or STAT1 phosphorylation in LPS/IFN-γ-stimulated cells. The nuclear transport of active NF-KB or STAT1 was also not affected by HSF1 inhibition. But HSF1 inhibition reduced the binding of NF-KB and STAT1 to their DNA elements. In addition, HSF1 inhibition reduced NF-KB and STAT1 bindings to iNOS promoter inside the LPS/IFN-γ-stimulated cells. Conclusions This preventing effect of HSF1 inhibition on iNOS mRNA transcription presents the necessary role of HSF1 in iNOS induction.展开更多
Aim To explore the role of transcription factor Foxp3 and the regulating effect of triptolide （TP） in the progression of myocardial hypertrophy in mice. Methods Fifty male mice were randomly divided into 5 groups, i...Aim To explore the role of transcription factor Foxp3 and the regulating effect of triptolide （TP） in the progression of myocardial hypertrophy in mice. Methods Fifty male mice were randomly divided into 5 groups, i. e., normal control group, myocardial hypertrophy model group and TP （10, 30, 90μg · kg^-1） treated groups. Myocardial hypertrophy was induced by isoprenaline （ISO） 5 mg kg^-1 once daily for 14 days. Triptolide was giv- en intraperitoneally once daily. Left ventricle tissue was subjected to HE staining and chemiluminescence technique to assess effects on hypertrophy, fibrosis and inflammation, quantitative assessment of hypertrophy regulatory genes were performed by qPCR and WB. Results After 14 days of treatment, myocardial expressions of Foxp3 and CD4 were significantly reduced in the model group compared with controls. The expression level of TGFβ1 in control group was lower, while that in model group increased obviously. TP could significantly lessen myocardial tissue damage, and reduce the heart index and left ventricular index. Compared with model group, TP （30, 90 μg · kg^-1 ） significantly increased myocardial expression ratio of α-MHC to β-MHC, reduced serumal levels of BNP and troponin I, elevated mRNA and protein expressions of Foxp3 and CD4 in myocardial tissue and reduced the protein expression of TGFβ1 by comparison of those in model group. Conclusion TP can effectively ameliorate myocardial damage and inhibit left ventricular remodeling through elevating the expression of CD4 and Foxp3 and decreasing that of TGF-β.展开更多
This paper presents an integrated approach towards solving the problem of "Gene Prediction".The "Gene Prediction" problem solving undergoes well defined stages starting with a DNA
Part 5＇ UTR region of pig SRPK1 gene was cloned by inverse PCR （I-PCR）, then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool （TFSEARCH） and 32 binding sites of tr...Part 5＇ UTR region of pig SRPK1 gene was cloned by inverse PCR （I-PCR）, then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool （TFSEARCH） and 32 binding sites of transcription with scores higher than 85 were getten, of which scores of Spl, MyoD, and HSF2 were over 90. Some of these binding sites of transcription factors were connected with promoters, but TATA-box, which was important to gene expression, hadn＇t been found in this region. By using PCR-SSCP method to search SNPs this part （5＇ UTR of SRPK1 in pig）, total of 40 Large White pigs were obtained as the research objects, but no polymorphism were found. Thus, 5＇ UTR of SRPK1 was speculated with a characteristic of high conservation, while it might have been directly or indirectly selected in commercial breeding. The paper provided a further feature of SRPK1 gene in molecular genetics.展开更多
Gene transcription mechanisms are critical control points for cell function and differentiation as well as disease pathology.It has remained difficult to target gene transcription mechanisms with small molecule drugs ...Gene transcription mechanisms are critical control points for cell function and differentiation as well as disease pathology.It has remained difficult to target gene transcription mechanisms with small molecule drugs due in part to the role of protein-protein interactions in transcription complexes.Rho A/C-GTPase regulation of the serum responsive transcription factor complex involving serum response factor(SRF)and myocardin-related transcription factor(MRTF)plays a key role in cancer and fibrotic mechanisms.In an attempt to disrupt this critical gene transcription mechanism,we undertook a high-throughput "pathway screen" using an SRE-Luciferase reporter which was activated by transient transfection of HEK293 cells with Ga13,an up-stream activator of Rho A and Rho C.The Rho/MRTF inhibitor tool compound CCG-1423 was identified in this screen.It and analogs such as CCG-203971have been used extensively to disrupt myofibroblast activation and tissue fibrosis as well as melanoma cell migration and metastasis.In the present study,we have used immobilized compounds and mass spectroscopy to identify the molecular target of the CCG-203971 series of anti-fibrotic and anti-metastatic agents.It is a poorly studied intranuclear protein that participates in gene transcription regulation by NF-κB and MRTF/SRF mechanisms.This dual mechanism rationalizes the strong efficacy of CCG-203971 and related compounds as anti-fibrotic and anti-metastatic agents.The identification of a molecular target also greatly facilitates future compound development through structure-based drug discovery and target biology evaluation.展开更多
Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible...Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible for transcriptional regulation and the model of their interplay in induction and repression will be presented. Using in vivo foot printing analysis of xylan-induced and glucose repressed mycelia, we detected three adjacent nucleotide sequences contacted by DNA-binding proteins. Protection within the inverted repeat of the Cre1 (SYGGRG) consensus sequence on the non coding strand under repressing conditions is in perfect agreement with the previously reported Cre1 dependent glucose repression of xyn1. Constitutive protein binding could be observed to a CCAAT-box and an inverted repeat of a 5′ GGCTAA 3′ sequence. EMSA with crude extracts from induced and repressed mycelia revealed that the latter motifs are sufficient for formation of the basal transcriptional complex under all conditions. The inverted repeat of GGCTAA closely resembles the consensus sequences of the cellulase and xylanase regulators Ace1, Ace2 and, Xyr1 (encoded by xyr1, cloned and characterised in this study) EMSA with heterologously expressed components of each factor and of the T. reesei Hap2/3/5 protein complex revealed that the basal transcriptional complex is formed by Xyr1 and the Hap2/3/5. Additionally to the Cre1 mediated carbon catabolite repression a yet unknown mechanism antagonizing induction of xyn1 expression could be elucidated. Latter occurs through competition of the repressor Ace1 and Xyr1 for the GGCTAA motif. In vivo proof for the relevance of identified motifs could be given through analysis of T. reesei transformants containing correspondingly mutated versions of the xyn1 promoter fused to the A. niger goxA gene. The results indicated that the basal as well as the induction level of xyn1 gene transcription is dependent on an interaction of Xyr1 with the GGCTAA motif while formation of the CCAAT-Hap2/3/5 complex slightly reduces induction. It can be concluded that mutations impairing protein binding in vitro lead to a loss of distinct regulatory functions in xyn1 gene expression in vivo. A respective model of gene regulation will be presented.展开更多
Cotton has enormous economic potential,providing high-quality protein,oil,and fibre.But the comprehensive utilization of cottonseed is limited by the presence of pigment gland and its inclusion.Pigment gland is a comm...Cotton has enormous economic potential,providing high-quality protein,oil,and fibre.But the comprehensive utilization of cottonseed is limited by the presence of pigment gland and its inclusion.Pigment gland is a common characteristic of Gossypium genus and its relatives,appearing as visible dark opaque dots in most tissues and organs of cotton plants.Secondary metabolites,such as gossypol,synthesized and stored in the cavities of pigment glands act as natural phytoalexins,but are toxic to humans and other monogastric animals.However,only a few cotton genes have been identified as being associated with pigment gland morphogenesis to date,and the developmental processes and regulatory mechanism involved in pigment gland formation remain largely unclear.Here,the research progress on the process of pigment gland morphogenesis and the genetic basis of cotton pigment glands is reviewed,for providing a theoretical basis for cultivating cotton with the ideal pigment gland trait.展开更多
Herpes simplex virus type 1 (HSV-1) naturallyestablishes latency in neurons of the nervous systemwith the concommitant expression of the latencyassociated transcripts (LATS) and the loss in
The human promyelocytic cell line HL-60 overexpresses the c-myc protooncogene. Plasmid pDACx carrying antisense human c-myc DNA and neo gene was introduced into HL-60 cells with lipofectin reagent. Upon DNA entering t...The human promyelocytic cell line HL-60 overexpresses the c-myc protooncogene. Plasmid pDACx carrying antisense human c-myc DNA and neo gene was introduced into HL-60 cells with lipofectin reagent. Upon DNA entering the tar-geted celis and expression of antisense transcripts to c-myc, C-MYC protein level, cell proliferation and colony-forming potentiality were all definitely inhibited.展开更多
While Upland cotton(Gossypium hirsutum L.) represents 95% of the world production,its genetic improvement is hindered by the shortage of effective genomic tools and resources.The
基金supported by the Natural Science Foundation of Hunan Province,China(2022JJ30886).
文摘Objective:Polycystic ovary syndrome(PCOS)is a common endocrine disorder that affects women’s health.This study aims to investigate gene and transcription factor(TF)expression differences between PCOS patients and healthy individuals using bioinformatics approaches,and to verify the function of key transcription factors,with the goal of providing new insights into the pathogenesis of PCOS.Methods:Differentially expressed genes(DEGs)and differentially expressed transcription factors(DETFs)between PCOS patients and controls were identified from the RNA sequencing dataset GSE168404 using bioinformatics methods.Functional enrichment analysis was performed using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases.The expression and function of core transcription factors were further validated in ovarian tissues of PCOS model mice and control mice using Western blotting and reverse transcription quantitative polymerase chain reaction(RTqPCR).Results:A total of 332 DEGs were identified between PCOS patients and controls,including 259 upregulated and 73 downregulated genes in the PCOS group.19 DETFs were further screened,of which 16 were upregulated and 3 were downregulated in PCOS.The upregulated DETFs(including TFCP2L1,DACH1,ESR2,AFF3,SMAD9,ZNF331,HOPX,ATOH8,HIF3α,DPF3,HOXC4,HES1,ID1,JDP2,SOX4,and ID3)were primarily associated with lipid metabolism,development,and cell adhesion.Protein and mRNA expression analysis in PCOS model mice revealed significantly decreased levels of hypoxia-inducible factor(HIF)1αand HIF2α,and significantly increased expression of HIF3αcompared to control mice(all P<0.001).Conclusion:Significant differences in gene and TF expression exist between PCOS patients and healthy individuals.HIF-3αmay play a crucial role in PCOS and could serve as a novel biomarker for diagnosis and a potential therapeutic target.
基金Supported by Cultivation of New Varieties of Genetically Modified Major Projects (2011ZX08004-005)Soybean Industry Technology System(CARS-04-PS08)
文摘Drought is one of the most important environmental constraints limiting plant growth, development and crop yield. Many drought-inducible genes have been identified by molecular and genomic analyses in Arabidopsis, rice and other crops. To better understand reaction mechanism of plant to drought tolerance, we mainly focused on introducing the research of transcription factors （TFs） in signal transduction and regulatory network of gene expression conferring drought. A TF could bind multiple target genes to increase one or more kinds of stress tolerance. Sometimes, several TFs might act together with a target gene. So drought-tolerance genes or TFs might respond to high-salinity, cold or other stresses. The crosstalk of multiple stresses signal pathways is a crucial aspect of understanding stress signaling.
文摘Abiotic stress including drought,lowtemperature,ABA and high salt is major factoraffecting the plant growth.Isolation andfunctional study of abiotic stress-related genewill be helpful to elucidate the signaltransduction mechanism of the target gene underabiotic stress during the growth of plant.Byusing gene-transfer technique,the target gene isincorporated into the plant to improve theadaptation of plant to abiotic stress.
基金supported by the Young Scientists Fund of the National Natural Science Foundation of China(32101797)Central Public-interest Scientific Institution Basal Research Fund(No.1610162023020)。
文摘Climate deterioration,water shortages,and abiotic stress are the main threats worldwide that seriously affect cotton growth,yield,and fiber quality.Therefore,research on improving cotton yield and tolerance to biotic and abiotic stresses is of great importance.The NAC proteins are crucial and plant-specific transcription factors(TFs)that are involved in cotton growth,development,and stress responses.The comprehensive utilization of cotton NAC TFs in the improvement of cotton varieties through novel biotechnological methods is feasible.Based on cotton genomic data,genome-wide identification and analyses have revealed potential functions of cotton NAC genes.Here,we comprehensively summarize the recent progress in understanding cotton NAC TFs roles in regulating responses to drought,salt,and Verticillium wilt-related stresses,as well as leaf senescence and the development of fibers,xylem,and glands.The detailed regulatory network of NAC proteins in cotton is also elucidated.Cotton NAC TFs directly bind to the promoters of genes associated with ABA biosynthesis and secondary cell-wall formation,participate in several biological processes by interacting with related proteins,and regulate the expression of downstream genes.Studies have shown that the overexpression of NAC TF genes in cotton and other model plants improve their drought or salt tolerance.This review elucidates the latest findings on the functions and regulation of cotton NAC proteins,broadens our understanding of cotton NAC TFs,and lays a fundamental foundation for further molecular breeding research in cotton.
基金Supported by National Natural Science Foundation of China(No.81370477No.81370918)+2 种基金Science and Technology Research Outstanding Youth Fund of Hebei College(No.Y2011117)Undergraduate Innovation Project(No.201410081061No.X2014026)
文摘AIM:Normalizing the results of real-time quantitative reverse transcription polymerase chain reaction(RT-q PCR)is essential for the accuracy of analysis.Commonly used approaches include input nucleic acid standardization(ΔCt method),normalization against a single internal reference gene(ΔΔCt method),and geometric averaging of multiple reference gene abundance using statistical software.We evaluated these approaches in the liver of db/db mice,a typical model of fatty liver disease.METHODS:Seven reference genes,β-actin(ACTB),eukaryotic initiation factor(e IF)5,glyceraldehyde-3-phosphate dehydrogenase(GAPDH),hydroxymethylbilane synthase(HMBS),hypoxanthineguanine phosphoribosyltransferase(HPRT)1,polymerase(RNA)II(DNA directed)polypeptide A(Polr2A)and ribosomal protein P〈0(RPLP〈0),were evaluated using software of ge Norm and Norm Finder.Hepatic lipogenesis genes,such as thyroid hormone-responsive protein(Thrsp),stearoyl-Co A desaturase(SCD)1,sterol regulatory element-binding protein(SREBP)1c and fatty acid synthase(FAS),were used as target genes of interest.RESULTS:The expression levels of all target genes and GAPDH were significantly elevated(P〈0.05)in db/db mouse livers by theΔCt method.ACTB and HMBS were the most stable genes calculated by the software of ge Norm.Norm Finder analysis indicated that ACTB was the most stable gene,and the best combination of 2 genes was GAPDH and RPLP〈0.Normalization against a single internal reference gene of ACTB or RPLP〈0,the geometric mean of ACTB and HMBS,or GAPDH and RPLP〈0 showed similar results that the expression levels of Thrsp,SCD1 and FAS,but not SREBP1 c increased(P〈0.05)in the liver of db/db mice.CONCLUSION:TheΔCt approach ensures a meaningful and biologically significant appraisal of gene expression.Use of the software like ge Norm or Norm Finder should be integrated withΔCt method.
基金Supported by the National Natural Science Foundation of China(31570189)。
文摘The myeloblastosis oncogenes(MYB)are important transcription factors that facilitate induction of variously develop-mental and stress responsive genes.They are hence,emerging as key players in improving stress tolerance of plants in response to several abiotic stresses.It was predicted that DfMYB2 contained an open reading frame(ORF)of 1621 bp coding 377 amino acid residues with molecular weight of 41.5 ku.It had 50 potential phosphorylation sites,44 potential N-glycosylation sites and one trans-membrane domain.In neighbor-joining(NJ)phylogenetic tree,DfMYB2 was close to the branch of angiosperms.The subcellular localization of DfMYB2 was in nucleus.The expressions of DfMYB2 increased gradually in the filament,prothalli and sporophyte.The results showed that DfMYB2 played a more and more important role in its growth and development.And the expression levels in rhizomes were significantly higher than those in roots and leaves.After abscisic acid(ABA)and PEG600 treatment,DfMYB2 showed a downward trend,followed by an upward trend.The expression of DfMYB2 was inhibited in a short time under drought stress,the strong induction of DfMYB2 expression by ABA indicated that it was possibly involved in abiotic stress responses in an ABA-dependent manner.
基金supported by the Major Research Plan of National Natural Science Foundation of China(NO.31690093)Creative Research Groups of China(31621005)the Agricultural Science and Technology Innovation Program Cooperation and Innovation Mission(CAAS-XTCX2016)
文摘Background:INDETERMINATE DOMAIN(IDD)transcription factors form one of the largest and most conserved gene families in plant kingdom and play important roles in various processes of plant growth and development,such as flower induction in term of flowering control.Till date,systematic and functional analysis of IDD genes remained infancy in cotton.Results:In this study,we identified total of 162 IDD genes from eight different plant species including 65 IDD genes in Gossypium hirsutum.Phylogenetic analysis divided IDDs genes into seven well distinct groups.The gene structures and conserved motifs of GhIDD genes depicted highly conserved exon-intron and protein motif distribution patterns.Gene duplication analysis revealed that among 142 orthologous gene pairs,54 pairs have been derived by segmental duplication events and four pairs by tandem duplication events.Further,Ka/Ks values of most of orthologous/paralogous gene pairs were less than one suggested the purifying selection pressure during evolution.Spatiotemporal expression pattern by qRT-PCR revealed that most of the investigated GhIDD genes showed higher transcript levels in ovule of seven days post anthesis,and upregulated response under the treatments of multiple abiotic stresses.Conclusions:Evolutionary analysis revealed that IDD gene family was highly conserved in plant during the rapid phase of evolution.Whole genome duplication,segmental as well as tandem duplication significantly contributed to the expansion of IDD gene family in upland cotton.Some distinct genes evolved into special subfamily and indicated potential role in the allotetraploidy Gossypium hisutum evolution and development High transcript levels of GhIDD genes in ovules illustrated their potential roles in seed and fiber development Further,upregulated responses of GhIDD genes under the treatments of various abiotic stresses suggested them as important genetic regulators to improve stress resistance in cotton breeding.
基金the Major Research Plan of National Natural Science Foundation of China(NO.31690093)the National Agricultural Science and Technology Innovation project for CAAS(CAAS-ASTIP-2016-ICR)the Central Level of the Scientific Research Institutes for Basic R&D Special Fund Business(Y2017PT51)。
文摘Background:Pectin is a key substance involved in cell wall development,and the galacturonosyltransferases(GAUTs)gene family is a critical participant in the pectin synthesis pathway.Systematic and comprehensive research on GAUTs has not been performed in cotton.Analysis of the evolution and expression patterns of the GAUT gene family in different cotton species is needed to in crease kno wledge of the functi on of pectin in cotto n fiber development.Results:In this study,we have identified 131 GAUT genes in the genomes of four Gossypium species(G.raimondii,G barbadense,G.hirsutum,and G.arboreum),and classified them as GAUT-A,GAUT-B and GAUT-C,which coding probable galacturonosyltransferases.Among them,the GAUT genes encode proteins GAUT1 to GAUT15.All GAUT proteins except for GAUT7 contai n a con served glycosyl transferase family 8 domain(H-DN-A-SW-S-V-H-T-F).The conserved sequence of GAUT7 is PLN(phospholamban)02769 domain.According to c/s-elemet analysis,GAUT genes transcript levels may be regulated by horm ones such as JA,GA,SA,ABA,Me-JA,and IA A.The evoluti on and transcription patterns of the GAUT gene family in different cotton species and the transcript levels in upland cotton lines with different fiber st「ength were analyzed.Peak transcript level of GhGAUT genes have been observed before 15 DPA.In the six materials with high fiber strength,the transcription of GhGAUT genes were concentrated from 10 to 15 DPA;while the highest transcript levels in low fiber st「ength materials were detected between 5 and 10 DPA.These results lays the foundation for future research on gene function during cotton fiber development.Conclusions:The GAUT gene family may affect cotton fiber development,including fiber elongation and fiber thickening.In the low strength fiber lines,GAUTs mainly participate in fiber elongation,whereas their major effect on cotton with high strength fiber is related to both elongation and thickening.
基金The National Natural Science Foundation of China(31621005,31530053,and 31671745)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences financially sponsored this research program.
文摘Background:Cotton is a valuable economic crop and the main significant source of natural fiber for textile industries globally.The effects of drought and salt stress pose a challenge to strong fiber and large-scale production due to the ever-changing climatic conditions.However,plants have evolved a number of survival strategies,among them is the induction of various stress-responsive genes such as the ribosomal protein large(RPL)gene.The RPL gene families encode critical proteins,which alleviate the effects of drought and salt stress in plants.In this study,comprehensive and functional analysis of the cotton RPL genes was carried out under drought and salt stresses.Results:Based on the genome-wide evaluation,26,8,and 5 proteins containing the RPL14B domain were identified in Gossypium hirsutum,G.raimondii,and G.arboreum,respectively.Furthermore,through bioinformatics analysis,key cis-regulatory elements related to RPL14B genes were discovered.The Myb binding sites(MBS),abscisic acid-responsive element(ABRE),CAAT-box,TATA box,TGACG-motif,and CGTCA-motif responsive to methyl jasmonate,as well as the TCA-motif responsive to salicylic acid,were identified.Expression analysis revealed a key gene,Gh_D01G0234(RPL14B),with significantly higher induction levels was further evaluated through a reverse genetic approach.The knockdown of Gh_D01G0234(RPL14B)significantly affected the performance of cotton seedlings under drought/salt stress conditions,as evidenced by a substantial reduction in various morphological and physiological traits.Moreover,the level of the antioxidant enzyme was significantly reduced in VIGS-plants,while oxidant enzyme levels increased significantly,as demonstrated by the higher malondialdehyde concentration level.Conclusion:The results revealed the potential role of the RPL14B gene in promoting the induction of antioxidant enzymes,which are key in oxidizing the various oxidants.The key pathways need to be investigated and even as we exploit these genes in the developing of more stress-resilient cotton germplasms.
文摘Aim Inducible nitric oxide synthase （iNOS） makes a great contribution to host defense and inflamma-tion. In many settings, lipopolysaccharide （LPS） induces iNOS expression through activation of the inhibitor of KB- α （IKB-α） -nuclear factor-KB （NF-KB） cascade, whereas interferon-γ （IFN-γ） acts through Janus kinase （ JAK）- signal transducer and activator of transcription 1 （ STAT1 ） signals. Heat shock factor 1 （ HSF1 ）, a major regulator of heat shock protein transcription, has been shown to regulate the production of pro-inflammatory cytokines such as tumor necrosis factor-α（TNF-α） and interleukin-6 （IL-6）. But it remains obscure whether and how HSF1 affects iNOS induction. Methods Western blot was used to measure the protein expression. The mRNA level was meas- ured by real time-PCR. Silence of HSF1 was achieved by small interfering RNA. Nitric oxide （NO） content and NF-KB binding activity were assayed by commercial kits. Chromatin immunoprecipitation （CHIP） was used to measure the binding activity of NF-KB and STAT1 to iNOS promoters. Results HSF1 inhibition or knockdown pre- vented the LPS- and/or IFN-γ-stimulated iNOS protein expression in cultured microglia. HSF1 inhibition blocked iNOS mRNA transcription. These inhibitory effects of HSF1 inhibition on iNOS expression were confirmed in brain tissues from endotoxemic mice. Further analysis showed that HSF1 inhibition had no effect on IKB-α degradation and NF-KB or STAT1 phosphorylation in LPS/IFN-γ-stimulated cells. The nuclear transport of active NF-KB or STAT1 was also not affected by HSF1 inhibition. But HSF1 inhibition reduced the binding of NF-KB and STAT1 to their DNA elements. In addition, HSF1 inhibition reduced NF-KB and STAT1 bindings to iNOS promoter inside the LPS/IFN-γ-stimulated cells. Conclusions This preventing effect of HSF1 inhibition on iNOS mRNA transcription presents the necessary role of HSF1 in iNOS induction.
文摘Aim To explore the role of transcription factor Foxp3 and the regulating effect of triptolide （TP） in the progression of myocardial hypertrophy in mice. Methods Fifty male mice were randomly divided into 5 groups, i. e., normal control group, myocardial hypertrophy model group and TP （10, 30, 90μg · kg^-1） treated groups. Myocardial hypertrophy was induced by isoprenaline （ISO） 5 mg kg^-1 once daily for 14 days. Triptolide was giv- en intraperitoneally once daily. Left ventricle tissue was subjected to HE staining and chemiluminescence technique to assess effects on hypertrophy, fibrosis and inflammation, quantitative assessment of hypertrophy regulatory genes were performed by qPCR and WB. Results After 14 days of treatment, myocardial expressions of Foxp3 and CD4 were significantly reduced in the model group compared with controls. The expression level of TGFβ1 in control group was lower, while that in model group increased obviously. TP could significantly lessen myocardial tissue damage, and reduce the heart index and left ventricular index. Compared with model group, TP （30, 90 μg · kg^-1 ） significantly increased myocardial expression ratio of α-MHC to β-MHC, reduced serumal levels of BNP and troponin I, elevated mRNA and protein expressions of Foxp3 and CD4 in myocardial tissue and reduced the protein expression of TGFβ1 by comparison of those in model group. Conclusion TP can effectively ameliorate myocardial damage and inhibit left ventricular remodeling through elevating the expression of CD4 and Foxp3 and decreasing that of TGF-β.
文摘This paper presents an integrated approach towards solving the problem of "Gene Prediction".The "Gene Prediction" problem solving undergoes well defined stages starting with a DNA
基金Supported by 11th Five-year Plan Key Projects of National Science and Technology (2008BADB2B01)
文摘Part 5＇ UTR region of pig SRPK1 gene was cloned by inverse PCR （I-PCR）, then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool （TFSEARCH） and 32 binding sites of transcription with scores higher than 85 were getten, of which scores of Spl, MyoD, and HSF2 were over 90. Some of these binding sites of transcription factors were connected with promoters, but TATA-box, which was important to gene expression, hadn＇t been found in this region. By using PCR-SSCP method to search SNPs this part （5＇ UTR of SRPK1 in pig）, total of 40 Large White pigs were obtained as the research objects, but no polymorphism were found. Thus, 5＇ UTR of SRPK1 was speculated with a characteristic of high conservation, while it might have been directly or indirectly selected in commercial breeding. The paper provided a further feature of SRPK1 gene in molecular genetics.
基金supported by NIH grants R01 AR066049(to SD Larsen) and R01 GM115459(to RRN)
文摘Gene transcription mechanisms are critical control points for cell function and differentiation as well as disease pathology.It has remained difficult to target gene transcription mechanisms with small molecule drugs due in part to the role of protein-protein interactions in transcription complexes.Rho A/C-GTPase regulation of the serum responsive transcription factor complex involving serum response factor(SRF)and myocardin-related transcription factor(MRTF)plays a key role in cancer and fibrotic mechanisms.In an attempt to disrupt this critical gene transcription mechanism,we undertook a high-throughput "pathway screen" using an SRE-Luciferase reporter which was activated by transient transfection of HEK293 cells with Ga13,an up-stream activator of Rho A and Rho C.The Rho/MRTF inhibitor tool compound CCG-1423 was identified in this screen.It and analogs such as CCG-203971have been used extensively to disrupt myofibroblast activation and tissue fibrosis as well as melanoma cell migration and metastasis.In the present study,we have used immobilized compounds and mass spectroscopy to identify the molecular target of the CCG-203971 series of anti-fibrotic and anti-metastatic agents.It is a poorly studied intranuclear protein that participates in gene transcription regulation by NF-κB and MRTF/SRF mechanisms.This dual mechanism rationalizes the strong efficacy of CCG-203971 and related compounds as anti-fibrotic and anti-metastatic agents.The identification of a molecular target also greatly facilitates future compound development through structure-based drug discovery and target biology evaluation.
文摘Xylanase 1 (Xyn1) is one of the two major representatives of the xylanase system of T. reesei; the mechanisms governing its expression were analysed throughout this study. All factors and regulatory motifs responsible for transcriptional regulation and the model of their interplay in induction and repression will be presented. Using in vivo foot printing analysis of xylan-induced and glucose repressed mycelia, we detected three adjacent nucleotide sequences contacted by DNA-binding proteins. Protection within the inverted repeat of the Cre1 (SYGGRG) consensus sequence on the non coding strand under repressing conditions is in perfect agreement with the previously reported Cre1 dependent glucose repression of xyn1. Constitutive protein binding could be observed to a CCAAT-box and an inverted repeat of a 5′ GGCTAA 3′ sequence. EMSA with crude extracts from induced and repressed mycelia revealed that the latter motifs are sufficient for formation of the basal transcriptional complex under all conditions. The inverted repeat of GGCTAA closely resembles the consensus sequences of the cellulase and xylanase regulators Ace1, Ace2 and, Xyr1 (encoded by xyr1, cloned and characterised in this study) EMSA with heterologously expressed components of each factor and of the T. reesei Hap2/3/5 protein complex revealed that the basal transcriptional complex is formed by Xyr1 and the Hap2/3/5. Additionally to the Cre1 mediated carbon catabolite repression a yet unknown mechanism antagonizing induction of xyn1 expression could be elucidated. Latter occurs through competition of the repressor Ace1 and Xyr1 for the GGCTAA motif. In vivo proof for the relevance of identified motifs could be given through analysis of T. reesei transformants containing correspondingly mutated versions of the xyn1 promoter fused to the A. niger goxA gene. The results indicated that the basal as well as the induction level of xyn1 gene transcription is dependent on an interaction of Xyr1 with the GGCTAA motif while formation of the CCAAT-Hap2/3/5 complex slightly reduces induction. It can be concluded that mutations impairing protein binding in vitro lead to a loss of distinct regulatory functions in xyn1 gene expression in vivo. A respective model of gene regulation will be presented.
基金National Key Technology R&D Program of China(2022YFF1001403)National Science Foundation of China(32101764).
文摘Cotton has enormous economic potential,providing high-quality protein,oil,and fibre.But the comprehensive utilization of cottonseed is limited by the presence of pigment gland and its inclusion.Pigment gland is a common characteristic of Gossypium genus and its relatives,appearing as visible dark opaque dots in most tissues and organs of cotton plants.Secondary metabolites,such as gossypol,synthesized and stored in the cavities of pigment glands act as natural phytoalexins,but are toxic to humans and other monogastric animals.However,only a few cotton genes have been identified as being associated with pigment gland morphogenesis to date,and the developmental processes and regulatory mechanism involved in pigment gland formation remain largely unclear.Here,the research progress on the process of pigment gland morphogenesis and the genetic basis of cotton pigment glands is reviewed,for providing a theoretical basis for cultivating cotton with the ideal pigment gland trait.
文摘Herpes simplex virus type 1 (HSV-1) naturallyestablishes latency in neurons of the nervous systemwith the concommitant expression of the latencyassociated transcripts (LATS) and the loss in
文摘The human promyelocytic cell line HL-60 overexpresses the c-myc protooncogene. Plasmid pDACx carrying antisense human c-myc DNA and neo gene was introduced into HL-60 cells with lipofectin reagent. Upon DNA entering the tar-geted celis and expression of antisense transcripts to c-myc, C-MYC protein level, cell proliferation and colony-forming potentiality were all definitely inhibited.
文摘While Upland cotton(Gossypium hirsutum L.) represents 95% of the world production,its genetic improvement is hindered by the shortage of effective genomic tools and resources.The
