国际免疫学杂志
國際免疫學雜誌
국제면역학잡지
INTERNATIONAL JOURNAL OF IMMUNOLOGY
2012年
4期
303-311
,共9页
HLA-DQA1%3’-UTR%MicroRNA%多态性%平衡选择
HLA-DQA1%3’-UTR%MicroRNA%多態性%平衡選擇
HLA-DQA1%3’-UTR%MicroRNA%다태성%평형선택
HLA-DQA1%3'-UTR%MicroRNA%Polymorphism%Balancing selection
目的 揭示HLA-DQAI基因3’-UTR区多态性的进化机制及其对基因表达的调控功能.方法 利用核苷酸变异率计算、系统发育树构建等群体遗传学分析方法将IMGT/HLA数据库中释放的20条HLA-DQAl全长序列多态性进行系统分析,揭示3’-UTR多态性的进化机制.利用miRanda vl.9软件提供的算法,对HLA-DQA1三种差异较大的3'UTR等位基因序列进行microRNA的靶位点预测,对分值较高的预测利用体外荧光细胞实验进行验证.结果 HLA-DQA1基因的3’-UTR具有极高的核苷酸变异率及跨物种多态性,表明其受到平衡选择的作用.预测出14种分值较高的microRNA-靶标复合 物,且14个microRNA与3种DQA1-3’-UTR等位基因靶标的结合自由能以及分值均有不同程度的差异.通过体外荧光表达实验证明,HLA-DQAl受到hsa-mir-658的负调控作用,且hsa-mir-658对DQAl-3’-UTR 3种等位基因特异性的结合对基因表达产生了影响.结论 HLA-DQAl基因表达水平受到microRNA的负调控,且HLA-DQA1 3’-UTR的多态性可影响与microRNA的结合而引起等位基因特异性表达.
目的 揭示HLA-DQAI基因3’-UTR區多態性的進化機製及其對基因錶達的調控功能.方法 利用覈苷痠變異率計算、繫統髮育樹構建等群體遺傳學分析方法將IMGT/HLA數據庫中釋放的20條HLA-DQAl全長序列多態性進行繫統分析,揭示3’-UTR多態性的進化機製.利用miRanda vl.9軟件提供的算法,對HLA-DQA1三種差異較大的3'UTR等位基因序列進行microRNA的靶位點預測,對分值較高的預測利用體外熒光細胞實驗進行驗證.結果 HLA-DQA1基因的3’-UTR具有極高的覈苷痠變異率及跨物種多態性,錶明其受到平衡選擇的作用.預測齣14種分值較高的microRNA-靶標複閤 物,且14箇microRNA與3種DQA1-3’-UTR等位基因靶標的結閤自由能以及分值均有不同程度的差異.通過體外熒光錶達實驗證明,HLA-DQAl受到hsa-mir-658的負調控作用,且hsa-mir-658對DQAl-3’-UTR 3種等位基因特異性的結閤對基因錶達產生瞭影響.結論 HLA-DQAl基因錶達水平受到microRNA的負調控,且HLA-DQA1 3’-UTR的多態性可影響與microRNA的結閤而引起等位基因特異性錶達.
목적 게시HLA-DQAI기인3’-UTR구다태성적진화궤제급기대기인표체적조공공능.방법 이용핵감산변이솔계산、계통발육수구건등군체유전학분석방법장IMGT/HLA수거고중석방적20조HLA-DQAl전장서렬다태성진행계통분석,게시3’-UTR다태성적진화궤제.이용miRanda vl.9연건제공적산법,대HLA-DQA1삼충차이교대적3'UTR등위기인서렬진행microRNA적파위점예측,대분치교고적예측이용체외형광세포실험진행험증.결과 HLA-DQA1기인적3’-UTR구유겁고적핵감산변이솔급과물충다태성,표명기수도평형선택적작용.예측출14충분치교고적microRNA-파표복합 물,차14개microRNA여3충DQA1-3’-UTR등위기인파표적결합자유능이급분치균유불동정도적차이.통과체외형광표체실험증명,HLA-DQAl수도hsa-mir-658적부조공작용,차hsa-mir-658대DQAl-3’-UTR 3충등위기인특이성적결합대기인표체산생료영향.결론 HLA-DQAl기인표체수평수도microRNA적부조공,차HLA-DQA1 3’-UTR적다태성가영향여microRNA적결합이인기등위기인특이성표체.
Objective To disclose the evolution mechanism of polymorphisms in.HLA-DQA1 3' UTR and their function on gene expression regulation.Methods The methods to study the population genetics,such as the nucleotide diversity computation,the phylogenetic tree construction,etc.,are used to analyze the polymorphism of 20 genomic full length sequences of HLA-DQAl,which are released in IMGT/HLA database.Miranda v1.9 is used to predict microRNA-targets for 3 different alleles of DQA1-3'-UTR.Top predictions are confirmed by Luciferase reporter analysis in vitro.Results We find high level of nucleotide diversity and trans-species polymorphism in DQA1-3'-UTR,suggesting balancing selection on this region.Fourteen top predictions of microRNA-targets are obtained.The score and free energy of microRNA-targets among 3 alleles are different.Luciferase reporter analysis indicates that HLA-DQA1 is under negative control by hsa-mir-658,and the allele-specific binding to microRNA influences the expression level of luciferase.Conclusions The expression of HLA-DQA1 is under negative regulation by microRNA.Moreover,the polymorphism of DQA1 3' -UTR may influence the binding ability with microRNA and produce the allele-specific expression.
