Abstract The dopamine transporter （DAT） is involved in the regulation of extracellular dopamine levels. A 40-bp variable-number tandem repeat （VNTR） polymorphism in the 3-untranslated region （3UTR） of the DAT ha...Abstract The dopamine transporter （DAT） is involved in the regulation of extracellular dopamine levels. A 40-bp variable-number tandem repeat （VNTR） polymorphism in the 3-untranslated region （3UTR） of the DAT has been reported to be associated with various phenotypes that are involved in the aberrant regulation of dopaminergic neu- rotransmission. In the present study, we found that miR- 137 and miR-491 caused a marked reduction of DAT expression, thereby influencing neuronal dopamine trans- port. Moreover, the regulation of miR-137 and miR-491 on this transport disappeared after the DAT was silenced. The miR-491 seed region that is located on the VNTR sequence in the 3＇UTR of the DAT and the regulatory effect of miR- 491 on the DAT depended on the VNTR copy-number. These data indicate that miR-137 and miR-491 regulate DAT expression and dopamine transport at the post- transcriptional level, suggesting that microRNA may be targeted for the treatment of diseases associated with DAT dysfunction.展开更多
The messenger RNA 3'-untranslated region(3'UTR)plays an important role in regulation of gene expres-sion on the posttranscriptional level. The 3'UTR con-trols gene expression via orchestrated interactionbe...The messenger RNA 3'-untranslated region(3'UTR)plays an important role in regulation of gene expres-sion on the posttranscriptional level. The 3'UTR con-trols gene expression via orchestrated interactionbetween the structural components of mRNAs(cis-ele-ment) and the specific trans-acting factors(RNA bind-ing proteins and non-coding RNAs). The crosstalk ofthese factors is based on the binding sequences and/or direct protein-protein interaction, or just functionalinteraction. Much new evidence that has accumulatedsupports the idea that several RNA binding factors canbind to common mRNA targets: to the non-overlappingbinding sites or to common sites in a competitive fash-ion. Various factors capable of binding to the sameRNA can cooperate or be antagonistic in their actions.The outcome of the collective function of all factorsbound to the same mRNA 3'UTR depends on manycircumstances, such as their expression levels, affinity to the binding sites, and localization in the cell, which can be controlled by various physiological conditions. Moreover, the functional and/or physical interactions of the factors binding to 3'UTR can change the character of their actions. These interactions vary during the cell cycle and in response to changing physiological condi-tions. Abnormal functioning of the factors can lead to disease. In this review we will discuss how alterations of these factors or their interaction can affect cancer development and promote or enhance the malignant phenotype of cancer cells. Understanding these altera-tions and their impact on 3'UTR-directed posttran-scriptional gene regulation will uncover promising new targets for therapeutic intervention and diagnostics. We will also discuss emerging new tools in cancer di-agnostics and therapy based on 3'UTR binding factors and approaches to improve them.展开更多
In a previous study,we found that long non-coding genes in Alzheimer’s disease(AD)are a result of endogenous gene disorders caused by the recruitment of microRNA(miRNA)and mRNA,and that miR-200a-3p and other represen...In a previous study,we found that long non-coding genes in Alzheimer’s disease(AD)are a result of endogenous gene disorders caused by the recruitment of microRNA(miRNA)and mRNA,and that miR-200a-3p and other representative miRNAs can mediate cognitive impairment and thus serve as new biomarkers for AD.In this study,we investigated the abnormal expression of miRNA and mRNA and the pathogenesis of AD at the epigenetic level.To this aim,we performed RNA sequencing and an integrative analysis of the cerebral cortex of the widely used amyloid precursor protein and presenilin-1 double transgenic mouse model of AD.Overall,129 mRNAs and 68 miRNAs were aberrantly expressed.Among these,eight down-regulated miRNAs and seven up-regulated miRNAs appeared as promising noninvasive biomarkers and therapeutic targets.The main enriched signaling pathways involved mitogen-activated kinase protein,phosphatidylinositol 3-kinase-protein kinase B,mechanistic target of rapamycin kinase,forkhead box O,and autophagy.An miRNA-mRNA network between dysregulated miRNAs and corresponding target genes connected with AD progression was also constructed.These miRNAs and mRNAs are potential biomarkers and therapeutic targets for new treatment strategies,early diagnosis,and prevention of AD.The present results provide a novel perspective on the role of miRNAs and mRNAs in AD.This study was approved by the Experimental Animal Care and Use Committee of Institute of Medicinal Biotechnology of Beijing,China(approval No.IMB-201909-D6)on September 6,2019.展开更多
目的研究非编码小RNA-1896（miRNA-1896）和miRNA-409—3p对细胞因子信号抑制蛋白-3（suppressors of cytokinesignaling-3，SOTS3）基因的调控作用，构建floes3基因3’端非翻译区（3’-untranslated re-gion，3’-UTR）野生型和突变型...目的研究非编码小RNA-1896（miRNA-1896）和miRNA-409—3p对细胞因子信号抑制蛋白-3（suppressors of cytokinesignaling-3，SOTS3）基因的调控作用，构建floes3基因3’端非翻译区（3’-untranslated re-gion，3’-UTR）野生型和突变型重组荧光素酶报告载体。方法以原代培养的小鼠星形胶质细胞总eDNA为模板，通过点突变和缺失突变的方式分别对SOCS33’-UTR序列中miRNA-1896和miRNA-409—3p种子区的结合位点CAGAGA（897—902位）和AACATT（1425—1430位）进行突变，并将野生型的3’-UTR序列与突变的3’-UTR序列分别插入到虫荧光素酶表达质粒pGL3-Promoter获得重组质粒，命名为pGL3-SOCS3-WT、pGL3-SOCS3-M1和pGL3-SOCS3-M2。将上述重组质粒分别与miRNA-1896和miRNA-409—3p共转染至HEK293T细胞中，测定虫荧光素酶的活性。结果酶切验证及测序结果表明，重组质粒pGL3-SOCS3-WT、pGL3-SOCS3-M1和pGL3-SOCS3-M2构建成功。与对照组相比，转染miRNA-1896和miRNA-409—3p均显著降低了pGL3-SOCS3-WT虫荧光素酶的活性，但对pGL3-SOCS3-M1和pGL3-SOCS3-M2荧光素酶的活性并无明显影响。结论成功构建了soc33’-UTR野生型及突变型的虫荧光素酶重组表达质粒，确认CA-GAGA（897—902位）和AACATr（1425—1430位）分别是miRNA-1896和miRNA-409—3p种子区与socs33’-UTR结合的关键位点。展开更多
基金supported by grants from the National Postdoctoral Science Foundation,China(2014M552219)the Natural Science Foundation of Guangdong Province,China(2015 A030313889,2015A030401013,2014A030313709,and 2014A030 313710)+1 种基金the Science and Technology Planning Project of Shenzhen Municipality,China(ZDSYS201504301045406,JCYJ20150403110 829621,JCYJ20150403091443301,JCYJ20140415162542975,JCYJ 20140415162338855,JCYJ20140828163634004,and JCYJ201206 16144352139)the Health and Family Planning Commission Project of Shenzhen Municipality,China(201401026)
文摘Abstract The dopamine transporter （DAT） is involved in the regulation of extracellular dopamine levels. A 40-bp variable-number tandem repeat （VNTR） polymorphism in the 3-untranslated region （3UTR） of the DAT has been reported to be associated with various phenotypes that are involved in the aberrant regulation of dopaminergic neu- rotransmission. In the present study, we found that miR- 137 and miR-491 caused a marked reduction of DAT expression, thereby influencing neuronal dopamine trans- port. Moreover, the regulation of miR-137 and miR-491 on this transport disappeared after the DAT was silenced. The miR-491 seed region that is located on the VNTR sequence in the 3＇UTR of the DAT and the regulatory effect of miR- 491 on the DAT depended on the VNTR copy-number. These data indicate that miR-137 and miR-491 regulate DAT expression and dopamine transport at the post- transcriptional level, suggesting that microRNA may be targeted for the treatment of diseases associated with DAT dysfunction.
文摘The messenger RNA 3'-untranslated region(3'UTR)plays an important role in regulation of gene expres-sion on the posttranscriptional level. The 3'UTR con-trols gene expression via orchestrated interactionbetween the structural components of mRNAs(cis-ele-ment) and the specific trans-acting factors(RNA bind-ing proteins and non-coding RNAs). The crosstalk ofthese factors is based on the binding sequences and/or direct protein-protein interaction, or just functionalinteraction. Much new evidence that has accumulatedsupports the idea that several RNA binding factors canbind to common mRNA targets: to the non-overlappingbinding sites or to common sites in a competitive fash-ion. Various factors capable of binding to the sameRNA can cooperate or be antagonistic in their actions.The outcome of the collective function of all factorsbound to the same mRNA 3'UTR depends on manycircumstances, such as their expression levels, affinity to the binding sites, and localization in the cell, which can be controlled by various physiological conditions. Moreover, the functional and/or physical interactions of the factors binding to 3'UTR can change the character of their actions. These interactions vary during the cell cycle and in response to changing physiological condi-tions. Abnormal functioning of the factors can lead to disease. In this review we will discuss how alterations of these factors or their interaction can affect cancer development and promote or enhance the malignant phenotype of cancer cells. Understanding these altera-tions and their impact on 3'UTR-directed posttran-scriptional gene regulation will uncover promising new targets for therapeutic intervention and diagnostics. We will also discuss emerging new tools in cancer di-agnostics and therapy based on 3'UTR binding factors and approaches to improve them.
基金This study was supported by the National Natural Science Foundation of China(General Program),No.81673411the United Fund Project of National Natural Science Foundation of China,No.U1803281+1 种基金Young Medical Talents Award Project of Chinese Academy of Medical Sciences,No.2018RC350013Chinese Academy of Medical Sciences Innovation Project for Medical Science,No.2017-I2M-1-016(all to RL).
文摘In a previous study,we found that long non-coding genes in Alzheimer’s disease(AD)are a result of endogenous gene disorders caused by the recruitment of microRNA(miRNA)and mRNA,and that miR-200a-3p and other representative miRNAs can mediate cognitive impairment and thus serve as new biomarkers for AD.In this study,we investigated the abnormal expression of miRNA and mRNA and the pathogenesis of AD at the epigenetic level.To this aim,we performed RNA sequencing and an integrative analysis of the cerebral cortex of the widely used amyloid precursor protein and presenilin-1 double transgenic mouse model of AD.Overall,129 mRNAs and 68 miRNAs were aberrantly expressed.Among these,eight down-regulated miRNAs and seven up-regulated miRNAs appeared as promising noninvasive biomarkers and therapeutic targets.The main enriched signaling pathways involved mitogen-activated kinase protein,phosphatidylinositol 3-kinase-protein kinase B,mechanistic target of rapamycin kinase,forkhead box O,and autophagy.An miRNA-mRNA network between dysregulated miRNAs and corresponding target genes connected with AD progression was also constructed.These miRNAs and mRNAs are potential biomarkers and therapeutic targets for new treatment strategies,early diagnosis,and prevention of AD.The present results provide a novel perspective on the role of miRNAs and mRNAs in AD.This study was approved by the Experimental Animal Care and Use Committee of Institute of Medicinal Biotechnology of Beijing,China(approval No.IMB-201909-D6)on September 6,2019.
文摘目的研究非编码小RNA-1896（miRNA-1896）和miRNA-409—3p对细胞因子信号抑制蛋白-3（suppressors of cytokinesignaling-3，SOTS3）基因的调控作用，构建floes3基因3’端非翻译区（3’-untranslated re-gion，3’-UTR）野生型和突变型重组荧光素酶报告载体。方法以原代培养的小鼠星形胶质细胞总eDNA为模板，通过点突变和缺失突变的方式分别对SOCS33’-UTR序列中miRNA-1896和miRNA-409—3p种子区的结合位点CAGAGA（897—902位）和AACATT（1425—1430位）进行突变，并将野生型的3’-UTR序列与突变的3’-UTR序列分别插入到虫荧光素酶表达质粒pGL3-Promoter获得重组质粒，命名为pGL3-SOCS3-WT、pGL3-SOCS3-M1和pGL3-SOCS3-M2。将上述重组质粒分别与miRNA-1896和miRNA-409—3p共转染至HEK293T细胞中，测定虫荧光素酶的活性。结果酶切验证及测序结果表明，重组质粒pGL3-SOCS3-WT、pGL3-SOCS3-M1和pGL3-SOCS3-M2构建成功。与对照组相比，转染miRNA-1896和miRNA-409—3p均显著降低了pGL3-SOCS3-WT虫荧光素酶的活性，但对pGL3-SOCS3-M1和pGL3-SOCS3-M2荧光素酶的活性并无明显影响。结论成功构建了soc33’-UTR野生型及突变型的虫荧光素酶重组表达质粒，确认CA-GAGA（897—902位）和AACATr（1425—1430位）分别是miRNA-1896和miRNA-409—3p种子区与socs33’-UTR结合的关键位点。
