DNMT1错义突变与神经管畸形易感性的相关性研究  

Association Study between DNMT1 Missense Mutations and Susceptibility to Neural Tube Defects

作　　者：徐畅 彭瑞 王红艳 卢磊[3] Xu Chang;Peng Rui;Wang Hongyan;Lu Lei(School of Life Sciences,Fudan University,Shanghai,200438;Obstetrics and Gynecology Hospital,Fudan University,Shanghai,200011;Institute of Metabolism&Integrative Biology,Fudan University,Shanghai,200433)

机构地区：[1]复旦大学生命科学学院,上海200438 [2]复旦大学附属妇产科医院,上海200011 [3]复旦大学代谢与整合生物学研究院,上海200433

出　　处：《基因组学与应用生物学》2021年第2期841-848,共8页Genomics and Applied Biology

基　　金：国家自然科学青年基金项目(NSFC81430005)资助。

摘　　要：DNMT1(DNA(cytosine-5)-methyltransferase 1)在神经管的闭合过程中发挥重要作用,已有研究表明,DNMT1敲除的小鼠会出现神经管畸形(neural tube defects,NTDs)的表型。为了进一步探索DNMT1基因错义突变与神经管畸形发生的相关性,本研究对来自中国汉族人群的101例神经管畸形患者血液样本和171例正常对照血液样本中提取的基因组DNA进行了全基因组测序。在这些病例的DNMT1基因上一共检测到了5个病例特有的错义突变,其中c.2626G>A和c.4381C>T突变在不同物种间高度保守,并且c.4381C>T突变在公共数据库中没有记录。蛋白免疫印迹实验显示c.290A>G,c.358G>C,c.410C>G,c.2626G>A和c.4381C>T这5个突变都使DNMT1蛋白表达量下调。免疫荧光实验显示这5个突变不影响DNMT1蛋白的核定位。我们的研究表明病例样本中DNMT1上鉴定到的5个病例特有的错义突变可能通过下调DNMT1的蛋白表达量而导致神经管畸形的发生。DNMT1(DNA(cytosine-5)-methyltransferase 1)plays an important role in neural tube closure,and previous studies have revealed that DNMT1-knockout mice displayed neural tube defects(NTDs)phenotypes.To further explore the association between DNMT1 missense mutations and NTDs,this study collected 101 NTD-patient blood samples and 171 healthy control blood samples from Chinese Han population and performed whole genome sequencing on genomic DNA collected from these samples.We detected 5 case specific missense mutations in DNMT1 gene among these samples,among which c.2626 G>A and c.4381 C>T nucleotides mutations were highly conserved between different species.Notably,c.4381 C>T variant hasn’t been reported in public database.Western blotting results showed that c.290 A>G,c.358 G>C,c.410 C>G,c.2626 G>A and c.4381 C>T mutations showed down-regulated DNMT1 protein levels.Immunofluorescence assays indicated that these five mutations didn’t affect the nuclear localization of DNMT1 protein.Our study revealed that these five case specific missense mutations possibly led to NTDs through down regulating DNMT1 protein levels.

关 键 词：神经管畸形(NTDs) DNMT1 错义突变 核定位 

分 类 号：R748[医药卫生—神经病学与精神病学]