人尿酸盐转运子1基因第3内含子＋11G＞A单核苷酸多态性功能研究  

Functional analysis of ＋ 11G 〉 A single nucleotide polymorphisms in intron 3 of human urate transporter 1 gene

作　　者：路杰[1] 王灿[1] 王瑶[1] 李长贵[1] 崔凌凌[1] 

机构地区：[1]青岛大学医学院附属医院痛风病重点实验室, 266003

出　　处：《中华内科杂志》2014年第7期550-554,共5页Chinese Journal of Internal Medicine

基　　金：国家自然科学基金（81070686、31371272）;山东省自然科学基金重点项目（zR2010HZ001）

摘　　要：目的 研究人尿酸盐转运子1（hURAT1）基因的第3内含子＋11G＞A的单核苷酸多态性（SNP）对该基因功能的影响.方法 以hURAT1的野生型和突变型重组质粒转染真核细胞后,比较mRNA表达情况;以hURAT1的cDNA为模板分别构建野生型、突变型和第5外显子缺失型质粒并合成mRNA,显微注射入斑马鱼胚胎卵黄中,观察不同hURAT1蛋白的亚细胞定位.结果 与野生型相比,hURAT1基因第3内含子＋11G＞A突变型重组质粒在细胞内同时表达两种hURAT1的mRNA转录剪接体.一种为野生型,另一种为第5外显子完全缺失型.亚细胞定位研究表明,hURAT1野生型蛋白在细胞膜上有明显的富集,细胞质中呈点状分布;hURAT1突变型和第5外显子缺失型蛋白均弥散地存在于细胞质中,细胞膜上无表达.结论 hURAT1基因第3内含子＋11G＞ASNP导致该基因mRNA转录出现可变剪接,产生第5外显子完全缺失的hURAT1剪接异构体,影响蛋白的亚细胞定位.Objective We reported previously that single nucleotide polymorphisms SNP） ＋ 11G 〉 A in intron 3 of the human urate transporter 1 （hURAT1） gene are associated with hyperuncaemia in Han Chinese.The aim of the present study was to evaluate the effect of the variants on hURAT1 function.Methods The wild-type,mutant-type hURAT1 and exon 5-null hURAT1 were constructed,and respectively microinjected into the zebrafish embryo yolks.The subcellular localization of different genotypes of hURAT1 was detected by confocal laser scanning microscope.Results Compared with wild type,the mutant recombinant plasmid transcribed two types of mRNA spliceosome,the wild type and the exon 5-null type.The hURAT1 wild type protein was prominent localized on cell membrane,while the mutant type and exon 5-null hURAT1 proteins were distributed uniform in the cytoplasm but not on the cell membrane.Conclusion The hURAT1 variant ＋ 11 G 〉 A resulted in an alternative splicing of hURAT1 mRNA-exon 5-null type.Its protein product exhibited a different subcellular localization compared with that of wild type.

关 键 词：高尿酸血症 多态性 单核苷酸 人尿酸盐转运子1 

分 类 号：R589.7[医药卫生—内分泌]