Functional characterization of missense variants in the creatine transporter gene (SLC6A8):: Improved diagnostic application
Functional characterization of missense variants in the creatine transporter gene (SLC6A8):: Improved diagnostic application
复制标题
DOI：
10.1002/humu.20532
复制
发表时间：
2007-09-01
期刊：
影响因子：
3.9
通讯作者：
Salomons, Gaja S.
中科院分区：
文献类型：
--
作者：
Rosenberg, Efraim H.;Munoz, Cristina Martinez;Salomons, Gaja S.
肌酸转运蛋白缺乏症是一种 X 连锁智力障碍，由肌酸转运蛋白基因 (SLC6A8) 突变引起。迄今为止，SLC6A8 基因的 20 个突变已被描述。我们开发了一种诊断方法来测试成纤维细胞中肌酸的吸收。此外，我们还扩展了检测方法来表征新型 SLC6A8 错义变体。通过定点诱变在 SLC6A8 cDNA 中引入了总共 13 个变体。所有变体均瞬时转染到 SLC6A8 缺陷的成纤维细胞中，并测试缺陷原代成纤维细胞中肌酸摄取的恢复情况。因此，我们证明了九个变体（p.Gly87Arg、p.Phe107del、p.Tyr317X、p.Asn336del、p.Cys337Trp、p.Ile347del、p.Pro390Leu、p.Arg391Trp和p.Pro554-Leu）是致病性突变，四个变体（p.Lys4Arg、 p.Gly26Arg、p.Met560Val 和 p.Val629Ile) 是非致病性的。本研究提供了一种改进的诊断工具来对未知意义的序列变异进行分类。
Creatine transporter deficiency is an X,linked mental retardation disorder caused by mutations in the creatine transporter gene (SLC6A8). So far, 20 mutations in the SLC6A8 gene have been described. We have developed a diagnostic assay to test creatine uptake in fibroblasts. Additionally, we expanded the assay to characterize novel SLC6A8 missense variants. A total of 13 variants were introduced in the SLC6A8 cDNA by site-directed mutagenesis. All variants were transiently transfected in SLC6A8-deficient fibroblasts and tested for restoration of creatine uptake in deficient primary fibroblasts. Thus, we proved that nine variants (p.Gly87Arg, p.Phe107del, p.Tyr317X, p.Asn336del, p.Cys337Trp, p.Ile347del, p.Pro390Leu, p.Arg391Trp, and p.Pro554-Leu) are pathogenic mutations and four variants (p.Lys4Arg, p.Gly26Arg, p.Met560Val, and p.Val629Ile) are nonpathogenic. The present study provides an improved diagnostic toot to classify sequence variants of unknown significance.