Nucleotide sequence of the partially deleted D4Z4 locus in a patient with FSHD identifies a putative gene within each 3.3 kb element
Nucleotide sequence of the partially deleted D4Z4 locus in a patient with FSHD identifies a putative gene within each 3.3 kb element
复制标题
DOI：
10.1016/s0378-1119(99)00267-x
复制
发表时间：
1999-08-05
期刊：
影响因子：
3.5
通讯作者：
Belayew, A
中科院分区：
文献类型：
--
作者：
Gabriëls, J;Beckers, MC;Belayew, A
面肩肱型肌营养不良症（FSHD）与染色体4 q35上的多态性D4 Z4基因座连锁。在未受影响的个体中，该基因座包含3.3 kb分散重复家族成员的10-100个串联拷贝。我们已经确定了一个13.5kb的EcoRI基因组片段的完整核苷酸序列，该片段包含一个患有FSHD的患者的受影响的D4 Z4基因座中仅存的两个3.3kb元件。序列分析表明，这两个3.3kb的重复序列是相同的。它们含有一个先前未检测到的推定启动子，具有TACAA而不是TATAA盒和GC盒。瞬时表达的荧光素酶报告基因融合到191 bp的这个启动子，表现出强烈的活性，在转染的人横纹肌肉瘤TE 671细胞，在TACAA或GC盒的突变的影响。此外，这些3.3 kb重复序列包括一个开放阅读框（ORF），该ORF起始于TACAA盒下游149 bp处，编码一个391个残基的具有两个同源结构域的蛋白（DUX 4）。兔网织红细胞裂解物中ORF的体外转录/翻译产生了两种S-35 CYs/ S-35 Met标记的产物，SDS-PAGE上的表观分子量分别为38和75 kDa，分别对应于DUX 4单体和二聚体。总之，我们提出部分缺失的D4 Z4基因座中的每个3.3kb元件可能包括编码双同源结构域蛋白的DUX 4基因。(C)1999 Elsevier Science B. V.保留所有权利。
Facioscapulohumeral muscular dystrophy (FSHD) is linked to the polymorphic D4Z4 locus on chromosome 4q35. In nonaffected individuals, this locus comprises 10-100 tandem copies of members of the 3.3 kb dispersed repeat family. Deletions leaving 1-8 such repeats have been associated with FSHD, for which no candidate gene has been identified.We have determined the complete nucleotide sequence of a 13.5 kb EcoRI genomic fragment comprising the only two 3.3 kb elements left in the affected D4Z4 locus of a patient with FSHD. Sequence analyses demonstrated that the two 3.3 kb repeats were identical. They contain a putative promoter that was not previously detected, with a TACAA instead of a TATAA box, and a GC box. Transient expression of a luciferase reporter gene fused to 191 bp of this promoter, demonstrated strong activity in transfected human rhabdomyosarcoma TE671 cells that was affected by mutations in the TACAA or GC box. In addition, these 3.3 kb repeats include an open reading frame (ORF) starting 149 bp downstream from the TACAA box and encoding a 391 residue protein with two homeodomains (DUX4). In-vitro transcription/translation of the ORF in a rabbit reticulocyte lysate yielded two S-35 CYs/ S-35 Met labeled products with apparent molecular weights of 38 and 75 kDa on SDS-PAGE, corresponding to the DUX4 monomer and dimer, respectively.In conclusion, we propose that each of the 3.3 kb elements in the partially deleted D4Z4 locus could include a DUX4 gene encoding a double homeodomain protein. (C) 1999 Elsevier Science B.V. All rights reserved.