Functional analysis of + 11G ＞ A single nucleotide polymorphisms in intron 3 of human urate transporter 1 gene / 中华内科杂志

ABSTRACT

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.