GM1 Gangliosidosis: Molecular Analysis of Nine Patients and Development of an RT-PCR Assay for GLB1 Gene Expression Profiling
GM1 Gangliosidosis: Molecular Analysis of Nine Patients and Development of an RT-PCR Assay for GLB1 Gene Expression Profiling
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DOI：
10.1002/humu.9475
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发表时间：
2007-02-01
期刊：
影响因子：
3.9
通讯作者：
Morrone, Amelia
中科院分区：
文献类型：
--
作者：
Caciotti, Anna;Donati, Maria Alice;Morrone, Amelia
人GLB 1基因产生两个可变剪接的转录本，编码溶酶体酶β-半乳糖苷酶（GLB 1）和弹性蛋白结合蛋白（EBP）。GLB 1基因座的突变，这是负责储存疾病GM 1神经节苷脂沉积症，可能会影响这两种蛋白质或GLB 1只。EBP，当受到影响时，有助于GM 1神经节苷脂沉积症患者的特定特征，如心肌病和结缔组织异常。在这里，我们报告了基于实时PCR的可靠和定量检测的发展，用于评估患者样本中GLB 1和EBP转录物的水平。我们还报告了9例GM 1神经节苷脂沉积症患者GLB 1基因突变的特征，以将遗传病变与mRNA水平和表型相关联。突变分析鉴定了4个新的（c.1835_1836delCC; p.Arg148Cys; c.1068+1G> T;和p.Pro549Leu），5个已知的（p.Arg59His; p.Arg201His; p.Gly123Arg; c.245+1G>A;和c.75+ 2 insT）突变和1个新的多态性（c.1233+ 8 T>C）。对患者表型的比较分析使GLB 1突变与特定临床表现之间的相关性更加彻底。在3名携带剪接缺陷的患者中，GLB 1和EBP mRNA水平均降低。这种准确、快速检测GLB 1基因可变剪接转录本的方法可应用于其他编码多种mRNA的致病溶酶体基因。(C)2007 Wiley-Liss，Inc.
The human GLB1 gene produces two alternatively spliced transcripts that encode the lysosomal enzyme beta-galactosidase (GLB1) and the elastin binding protein (EBP). Mutations at the GLB1 locus, which are responsible for the storage disorder GM1 gangliosidosis, may affect either both proteins or GLB1 only. The EBP, when affected, contributes to specific features of GM1 gangliosidosis patients, such as cardiomyopathy and connective-tissue abnormalities. Here we report the development of reliable and quantitative assays based on real-time PCR for assessing the levels of GLB1 and EBP transcripts in patients' samples. We also report the characterisation of GLB1 gene mutations in nine GM1 gangliosidosis patients in order to correlate the genetic lesions with mRNA levels and phenotypes. Mutation analysis identified four new (c.1835_1836delCC; p.Arg148Cys; c.1068+1G> T; and p.Pro549Leu), five known (p.Arg59His; p.Arg201His; p.Gly123Arg; c.245+1G>A; and c.75+2insT) mutations and one new polymorphism (c.1233+8T>C). Comparative analysis of the patients' phenotypes enabled a more thorough correlation between GLB1 mutations and specific clinical manifestations. GLB1 and EBP mRNA levels were both reduced in three patients carrying the splicing defects. The accurate and fast method for the detection of alternatively spliced transcripts of the GLB1 gene could be applied to other disease-causing lysosomal genes that encode multiple mRNAs. (C) 2007 Wiley-Liss, Inc.