突触黏附分子编码基因NRXN2α启动子的功能分析  

Functional Analysis of the Promoter of NRXN2α Gene Coding Synaptic Adhesion Molecule

作　　者：周家红 郭印 孟莎莎[1] 周卫辉[1] ZHOU Jiahong;GUO Yin;MENG Shasha;ZHOU Weihui(Department of Pediatric Research Institute,Ministry of Education Key Laboratory of Child Development and Disorders,Chongqing Key Laboratorly ofTranslational Medical Research in Cognitive Development and Learning and Memory Disorders,National Clinical ResearchCenter for Child Health and Disorders,China International Science and Technology Cooperation base of Child development and Critical Disorders,Children's Hospital of Chongqing Medical University,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院儿科研究所,儿童发育疾病研究教育部重点实验室,认知发育与学习记忆障碍转化医学重庆市重点实验室,国家儿童健康与疾病临床医学研究中心,儿童发育重大疾病国家国际科技合作基地,重庆400014

出　　处：《中国细胞生物学学报》2020年第6期997-1007,共11页Chinese Journal of Cell Biology

基　　金：国家重点基础研究发展计划(973项目)(批准号:2012CB517903);国家自然科学基金(批准号:81070269、81571388)资助的课题。

摘　　要：Neurexins是突触前膜细胞黏附分子,参与神经元和脑回路的跨突触信号传递,其基因表达水平的调控密切影响着突触可塑性和特异性。为分析NRXN2α基因转录水平的调控,该研究构建了含有NRXN2α基因5′端不同区域的荧光素酶报告基因质粒,转染HEK293和U-87 MG细胞并检测双荧光素酶报告基因的表达水平,该研究确定了具有较强转录活性的NRXN2α基因启动子区域–911/+60(转录起始位点为+1),并鉴定了具有基础转录活性的最小启动子区域–58/+60,以及4个正向调控功能区域–911/–629、–109/–76、–49/–17、–17/+60,和1个负向调控功能区域+146/+238。最小启动子区域序列分析鉴定了1个典型的核心启动子元件initiator(Inr)。总之,该研究首次克隆了NRXN2α基因的启动子区域并进行了功能分析,为进一步阐明转录调控机制和加深对突触可塑性的理解奠定了基础。Neurexins are a family of presynaptic adhesion proteins which play critical roles in overall trans-synaptic signaling network in neurons and circuits.Regulation of gene expression in transcriptional and post-transcriptional levels affects synaptic plasticity and synaptic specificity.To investigate the transcriptional regulation of NRXN2αgene,luciferase reporter plasmids containing different regions of 5′end of NRXN2αgene were constructed and transfected into HEK293 and U-87 MG cells.Then,the promoter activity was determined by the luciferase activity assay of reporter genes.Here,the promoter region of human NRXN2αgene(–911/+60,+1 being the transcription start site)was defined.Then,further study identified the minimal promoter region of NRXN2α gene(–58/+60),which was sufficient to initiate a basal transcription.Moreover,four regions with positive regulatory function(–911/–629,109/–76,–49/–17,and–17/+60)and one region with negative regulatory function(+146/+238)for transcription of NRXN2αgene were defined.In addition,the typical Inr(initiator)motif in the minimal promoter was characterized.Collectively,for the first time,the promoter region of NRXN2αgene is characterized,and its function in regulating gene expression is analyzed,which lays a foundation for further elucidating its transcriptional regulation mechanism.

关 键 词：NRXN2α 启动子 转录 突触 

分 类 号：Q75[生物学—分子生物学]