AIM TO uncover the roles of tumor-promoting gene ZEB1 in aerobic glycolysis regulation and shed light on the underlying molecular mechanism.METHODS Endogenous zinc finger E-box binding homeobox-1 （ZEB1） was silenced...AIM TO uncover the roles of tumor-promoting gene ZEB1 in aerobic glycolysis regulation and shed light on the underlying molecular mechanism.METHODS Endogenous zinc finger E-box binding homeobox-1 （ZEB1） was silenced using a and the impact of ZEB1 and lentivirus-mediated method, methyI-CpG binding domain protein 1 （MBD1） on aerobic glycolysis was measured using seahorse cellular flux analyzers, reactive oxygen species quantification, and mitochondrial membrane potential measurement. The interaction between ZEB1 and MBD1 was assessed by co-immunoprecipitation and immunofluorescence assays. The impact of ZEB1 and MBD1 interaction on sirtuin 3 （SIRT3） expression was confirmed by quantitative polymerase chain reaction, western blotting, and dual-luciferase and chromatinimmunoprecipitation assays.RESULTS ZEB1 was a positive regulator of aerobic glycolysis in pancreatic cancer. ZEB1 transcriptionally silenced expression of SIRT3, a mitochondrial-localized tumor suppressor, through interaction with MBD1.CONCLUSION ZEB1 silenced SIRT3 expression via interaction with MBD1 to promote aerobic glycolysis in pancreatic cancer.展开更多
成簇的规律间隔的短回文重复序列及其相关蛋白9〔clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated protein 9(Cas9),CRISPR/Cas9〕是一种新兴的基因编辑技术,与以前的三大基因编辑技术——归巢核...成簇的规律间隔的短回文重复序列及其相关蛋白9〔clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated protein 9(Cas9),CRISPR/Cas9〕是一种新兴的基因编辑技术,与以前的三大基因编辑技术——归巢核酸内切酶、锌指核酸酶和转录激活因子样效应物核酸酶技术相比,其在靶向特异性、操作简便性、治疗彻底性、应用广泛性等方面具有更大的优势和发展潜力。艾滋病、乙型肝炎、疟疾等感染性疾病的治疗一直是医学上的重大难题,科学家正努力尝试利用CRISPR/Cas9技术解决这些医学难题。本文主要综述了CRISPR/Cas9技术在这些感染性疾病中应用的研究进展。展开更多
Previously, we found 123 transcription factors（TFs） as candidate regulators of secondary cell wall（SCW）formation in rice by using phylogenetic and co-expression network analyses. Among them, we examined in this wo...Previously, we found 123 transcription factors（TFs） as candidate regulators of secondary cell wall（SCW）formation in rice by using phylogenetic and co-expression network analyses. Among them, we examined in this work the role of OsIDD_2, a zinc finger and indeterminate domain（IDD） family TF. Its overexpressors showed dwarfism, fragile leaves, and decreased lignin content, which are typical phenotypes of plants defective in SCW formation, whereas its knockout plants showed slightly increased lignin content.The RNA-seq and quantitative reverse transcription polymerase chain reaction analyses confirmed that some lignin biosynthetic genes were downregulated in the OsIDD_2-overexpressing plants, and revealed the same case for other genes involved in cellulose synthesis and sucrose metabolism. The transient expression assay using rice protoplasts revealed that OsIDD_2 negatively regulates the transcription of genes involved in lignin biosynthesis, cinnamyl alcohol dehydrogenase 2 and 3（CAD_2 and 3）, and sucrose metabolism, sucrose synthase 5（SUS_5）, whereas an Alpha Screen assay, which can detect the interaction between TFs and their target DNA sequences, directly confirmed the interaction between OsIDD_2 and the target sequences located in the promoter regions of CAD_2 and CAD_3. Based on these observations, we conclude that OsIDD_2 is negatively involved in SCW formation and other biological events by downregulating its target genes.展开更多
GENETIC algorithms are very efficient search mechanisms. The application of genetic algo-rithms in the protein structure study provides not only a convenience algorithm for the searchof the lowest energy status (accor...GENETIC algorithms are very efficient search mechanisms. The application of genetic algo-rithms in the protein structure study provides not only a convenience algorithm for the searchof the lowest energy status (according to the thermodynamics theory, the natural protein展开更多
The second zinc finger fragment of Sp1 (Spl-ZF2), its mutant (Spl-ZF2/HT. E20→H, R23→T), and two mimic analogues (ZF20 and ZF15) were synthesized by stepwise solid phase technique. The CD spectra and UV-visible spec...The second zinc finger fragment of Sp1 (Spl-ZF2), its mutant (Spl-ZF2/HT. E20→H, R23→T), and two mimic analogues (ZF20 and ZF15) were synthesized by stepwise solid phase technique. The CD spectra and UV-visible spectrum with CoCl2 indicated that the formation of zinc finger structure was affected not only by the hy-drophobic amino acids but also by the change of the distance between Cys and His. Gel-retardation electrophoresis as-says indicated that the Grlu and Arg residues are very important for recognition. A single zinc finger like Spl-ZF2 isable to bind DNA sequence specifically.展开更多
目的研究毗邻锌指结构域的溴结构域蛋白2A(bromodomain adjacent to zinc finger domain protein 2,BAZ2A)促进子宫颈癌和肝癌发展的共同机制。方法通过转录组测序获得子宫颈癌组和肝癌组的转录组数据。应用R语言的“limma”包分别筛选...目的研究毗邻锌指结构域的溴结构域蛋白2A(bromodomain adjacent to zinc finger domain protein 2,BAZ2A)促进子宫颈癌和肝癌发展的共同机制。方法通过转录组测序获得子宫颈癌组和肝癌组的转录组数据。应用R语言的“limma”包分别筛选子宫颈癌组DEGs和肝癌组DEGs,并取交集获得其共有DEGs。通过“ggplot2”和“clusterProfiler”包对DEGs进行GO和KEGG功能注释分析。应用STRING数据库在线工具构建子宫颈癌DEGs、肝癌DEGs和共有DEGs的PPI网络分析图。使用Cytoscape软件对PPI网络分析图进行进一步处理,鉴定出核心基因。结果对子宫颈癌DEGs、肝癌DEGs和共有DEGs分别进行KEGG富集,三者共有的通路涉及细胞凋亡、抗原加工与呈递、类固醇生物合成。对子宫颈癌DEGs、肝癌DEGs和共有DEGs分别进行GO富集,前两者共有的生物过程(biological process,BP)条目涉及凋亡信号通路、免疫反应、生物黏附,三者共有的BP条目为细胞-底物黏附。子宫颈癌DEGs的核心基因为EP300。EP300对癌症细胞凋亡、迁移有调控作用。肝癌DEGs的核心基因为HSP90AB1。HSP90AB1可介导细胞程序性死亡、炎症和自身免疫、迁移等过程。共有DEGs的核心基因为RPS3。RPS3是一种核糖体蛋白,可通过影响核糖体的生物发生而影响癌细胞的生长、增殖和转移。结论BAZ2A可通过调节细胞凋亡、免疫反应、细胞运动、迁移而影响子宫颈癌、肝癌的发展,这为癌症的靶向治疗提供了新思路。展开更多
The stress-associated protein SAP12 belongs to the stress-associated protein （SAP） family with 14 members in Arabidopsis thaliana. SAP12 contains two AN1 zinc fingers and was identified in diagonal 2D redox SDS-PAGE...The stress-associated protein SAP12 belongs to the stress-associated protein （SAP） family with 14 members in Arabidopsis thaliana. SAP12 contains two AN1 zinc fingers and was identified in diagonal 2D redox SDS-PAGE as a protein undergoing major redox-dependent conformational changes. Its transcript was strongly induced under cold and salt stress in a time-dependent manner similar to SAP10, with high levels after 6 h and decreasing levels after 24 and 48 h. The tran- script regulation resembled those of the stress marker peroxiredoxin PrxllD at 24 and 48 h. Recombinant SAP12 protein showed redox-dependent changes in quaternary structure as visualized by altered electrophoretic mobility in non-reducing SDS polyacrylamide gel electrophoresis. The oxidized oligomer was reduced by high dithiothreitol concentrations, and also by E. coli thioredoxin TrxA with low dithiothreitol （DTF） concentrations or NADPH plus NADPH-dependent thioredoxin reductase. From Western blots, the SAP12 protein amount was estimated to be in the range of 0.5 ngμg^-1 leaf protein. SAP12 protein decreased under salt and cold stress. These data suggest a redox state-linked function of SAP12 in plant cells particularly under cold and salt stress.展开更多
基金the National Science Fund for Distinguished Young Scholars of China,No.81625016the National Science Foundation of China,No.81502031 and No.81772555+1 种基金Shanghai Municipal Commission of Health and Family Planning Grant,No.20154Y0090Youth Research Foundation of Shanghai Municipal Commission of Health and Family Planning,No.Z0124Y074
文摘AIM TO uncover the roles of tumor-promoting gene ZEB1 in aerobic glycolysis regulation and shed light on the underlying molecular mechanism.METHODS Endogenous zinc finger E-box binding homeobox-1 （ZEB1） was silenced using a and the impact of ZEB1 and lentivirus-mediated method, methyI-CpG binding domain protein 1 （MBD1） on aerobic glycolysis was measured using seahorse cellular flux analyzers, reactive oxygen species quantification, and mitochondrial membrane potential measurement. The interaction between ZEB1 and MBD1 was assessed by co-immunoprecipitation and immunofluorescence assays. The impact of ZEB1 and MBD1 interaction on sirtuin 3 （SIRT3） expression was confirmed by quantitative polymerase chain reaction, western blotting, and dual-luciferase and chromatinimmunoprecipitation assays.RESULTS ZEB1 was a positive regulator of aerobic glycolysis in pancreatic cancer. ZEB1 transcriptionally silenced expression of SIRT3, a mitochondrial-localized tumor suppressor, through interaction with MBD1.CONCLUSION ZEB1 silenced SIRT3 expression via interaction with MBD1 to promote aerobic glycolysis in pancreatic cancer.
文摘成簇的规律间隔的短回文重复序列及其相关蛋白9〔clustered regularly interspaced short palindromic repeat(CRISPR)/CRISPR-associated protein 9(Cas9),CRISPR/Cas9〕是一种新兴的基因编辑技术,与以前的三大基因编辑技术——归巢核酸内切酶、锌指核酸酶和转录激活因子样效应物核酸酶技术相比,其在靶向特异性、操作简便性、治疗彻底性、应用广泛性等方面具有更大的优势和发展潜力。艾滋病、乙型肝炎、疟疾等感染性疾病的治疗一直是医学上的重大难题,科学家正努力尝试利用CRISPR/Cas9技术解决这些医学难题。本文主要综述了CRISPR/Cas9技术在这些感染性疾病中应用的研究进展。
基金supported by grants from Grant-in-Aid for JSPS Research Fellow (JP15J03980 to H.Y.)Series of single-year grants (26-1393 to K.H.)+1 种基金Grants-in-Aid for Scientific Research on Innovative Areas (No.3806 to M.M. and M. U.-T.)the Grant-in-Aid for Scientific Research (A) (JP26252001 to M.M.) and (B) (JP16H04907 to M. U.-T.)
文摘Previously, we found 123 transcription factors（TFs） as candidate regulators of secondary cell wall（SCW）formation in rice by using phylogenetic and co-expression network analyses. Among them, we examined in this work the role of OsIDD_2, a zinc finger and indeterminate domain（IDD） family TF. Its overexpressors showed dwarfism, fragile leaves, and decreased lignin content, which are typical phenotypes of plants defective in SCW formation, whereas its knockout plants showed slightly increased lignin content.The RNA-seq and quantitative reverse transcription polymerase chain reaction analyses confirmed that some lignin biosynthetic genes were downregulated in the OsIDD_2-overexpressing plants, and revealed the same case for other genes involved in cellulose synthesis and sucrose metabolism. The transient expression assay using rice protoplasts revealed that OsIDD_2 negatively regulates the transcription of genes involved in lignin biosynthesis, cinnamyl alcohol dehydrogenase 2 and 3（CAD_2 and 3）, and sucrose metabolism, sucrose synthase 5（SUS_5）, whereas an Alpha Screen assay, which can detect the interaction between TFs and their target DNA sequences, directly confirmed the interaction between OsIDD_2 and the target sequences located in the promoter regions of CAD_2 and CAD_3. Based on these observations, we conclude that OsIDD_2 is negatively involved in SCW formation and other biological events by downregulating its target genes.
文摘GENETIC algorithms are very efficient search mechanisms. The application of genetic algo-rithms in the protein structure study provides not only a convenience algorithm for the searchof the lowest energy status (according to the thermodynamics theory, the natural protein
基金Project supported by the Fok Ying Tung Education Foundation, the National Natural Science Foundation of China, and the State Education Commission of China.
文摘The second zinc finger fragment of Sp1 (Spl-ZF2), its mutant (Spl-ZF2/HT. E20→H, R23→T), and two mimic analogues (ZF20 and ZF15) were synthesized by stepwise solid phase technique. The CD spectra and UV-visible spectrum with CoCl2 indicated that the formation of zinc finger structure was affected not only by the hy-drophobic amino acids but also by the change of the distance between Cys and His. Gel-retardation electrophoresis as-says indicated that the Grlu and Arg residues are very important for recognition. A single zinc finger like Spl-ZF2 isable to bind DNA sequence specifically.
文摘目的研究毗邻锌指结构域的溴结构域蛋白2A(bromodomain adjacent to zinc finger domain protein 2,BAZ2A)促进子宫颈癌和肝癌发展的共同机制。方法通过转录组测序获得子宫颈癌组和肝癌组的转录组数据。应用R语言的“limma”包分别筛选子宫颈癌组DEGs和肝癌组DEGs,并取交集获得其共有DEGs。通过“ggplot2”和“clusterProfiler”包对DEGs进行GO和KEGG功能注释分析。应用STRING数据库在线工具构建子宫颈癌DEGs、肝癌DEGs和共有DEGs的PPI网络分析图。使用Cytoscape软件对PPI网络分析图进行进一步处理,鉴定出核心基因。结果对子宫颈癌DEGs、肝癌DEGs和共有DEGs分别进行KEGG富集,三者共有的通路涉及细胞凋亡、抗原加工与呈递、类固醇生物合成。对子宫颈癌DEGs、肝癌DEGs和共有DEGs分别进行GO富集,前两者共有的生物过程(biological process,BP)条目涉及凋亡信号通路、免疫反应、生物黏附,三者共有的BP条目为细胞-底物黏附。子宫颈癌DEGs的核心基因为EP300。EP300对癌症细胞凋亡、迁移有调控作用。肝癌DEGs的核心基因为HSP90AB1。HSP90AB1可介导细胞程序性死亡、炎症和自身免疫、迁移等过程。共有DEGs的核心基因为RPS3。RPS3是一种核糖体蛋白,可通过影响核糖体的生物发生而影响癌细胞的生长、增殖和转移。结论BAZ2A可通过调节细胞凋亡、免疫反应、细胞运动、迁移而影响子宫颈癌、肝癌的发展,这为癌症的靶向治疗提供了新思路。
文摘The stress-associated protein SAP12 belongs to the stress-associated protein （SAP） family with 14 members in Arabidopsis thaliana. SAP12 contains two AN1 zinc fingers and was identified in diagonal 2D redox SDS-PAGE as a protein undergoing major redox-dependent conformational changes. Its transcript was strongly induced under cold and salt stress in a time-dependent manner similar to SAP10, with high levels after 6 h and decreasing levels after 24 and 48 h. The tran- script regulation resembled those of the stress marker peroxiredoxin PrxllD at 24 and 48 h. Recombinant SAP12 protein showed redox-dependent changes in quaternary structure as visualized by altered electrophoretic mobility in non-reducing SDS polyacrylamide gel electrophoresis. The oxidized oligomer was reduced by high dithiothreitol concentrations, and also by E. coli thioredoxin TrxA with low dithiothreitol （DTF） concentrations or NADPH plus NADPH-dependent thioredoxin reductase. From Western blots, the SAP12 protein amount was estimated to be in the range of 0.5 ngμg^-1 leaf protein. SAP12 protein decreased under salt and cold stress. These data suggest a redox state-linked function of SAP12 in plant cells particularly under cold and salt stress.
