Functional Analysis of a Novel FOXL2 Indel Mutation in Chinese Families with Blepharophimosis-Ptosis-Epicanthus Inversus Syndrome Type I.
Functional Analysis of a Novel FOXL2 Indel Mutation in Chinese Families with Blepharophimosis-Ptosis-Epicanthus Inversus Syndrome Type I.
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中国I型睑裂-下垂-内眦赘皮综合征家系中新型FOXL2插入缺失突变的功能分析
DOI：
10.7150/ijbs.19532
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发表时间：
2017
影响因子：
9.2
通讯作者：
Fan X
中科院分区：
文献类型：
--
作者：
Chai P;Li F;Fan J;Jia R;Zhang H;Fan X
背景：小睑裂-上睑下垂-内眦赘皮综合征（BPES）是一种发病率低的常染色体显性遗传病。叉头框L2（FOXL 2）基因中的插入缺失突变导致两种类型的BPES，其通过卵巢早衰（POF）的存在（I型）或不存在（II型）来区分。本研究的目的是确定中国BPES家系中FOXL 2基因可能的缺失，并阐明其与POF的关系。方法：收集一个四代常染色体显性遗传的BPES家系。收集所有受影响患者的外周静脉血，并从白细胞中提取基因组DNA。采用3对重叠引物，通过聚合酶链反应（PCR）扩增FOXL 2基因的全编码序列及其附近的5'非翻译区（UTR）和3'非翻译区（3 'UTR），并进行测序分析。使用SeqMan软件分析测序结果。基于患者的临床表现和对所鉴定的indel突变的分析，我们发现该突变干扰了FOXL 2和星星基因之间的相互作用。此外，通过亚细胞定位和功能研究，我们观察到显着的错误定位的突变蛋白质，突变蛋白质被发现在细胞质中，而野生型蛋白质被发现在细胞核中。通过转录活性测定、定量实时PCR和电泳迁移率变动测定证实功能丧失。结果如下：所有患者的临床特征均为BPES I型，包括小睑裂、上睑下垂、内眦赘皮和内眦赘皮伴POF。一种新的FOXL 2杂合indel突变，c.19_95del，一个77 bp的缺失，破坏FOXL 2蛋白质结构，在所有受影响的家庭成员中被确定。此外，该indel突变通过破坏FOXL 2蛋白结合星星启动子并作为该基因的阻遏物的能力而显著增加星星mRNA表达。结论：在中国BPES家系中发现一种新的FOXL 2插入缺失突变。我们的研究结果扩展了已知FOXL 2突变的范围，并为FOXL 2蛋白的结构-功能关系提供了额外的见解。此外，这种新的突变导致FOXL 2作为转录因子的功能障碍，阻断其与星星基因启动子区结合的能力，导致受影响患者的POF。
Background: Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is an autosomal dominant disease with a low incidence rate. Indel mutations in the forkhead box L2 (FOXL2) gene cause two types of BPES that are distinguished by the presence (type I) or absence (type II) of premature ovarian failure (POF). The purpose of this study was to identify a possible deletion in FOXL2 in Chinese families with BPES and to clarify its relationship with POF. Methods: An autosomal dominant Chinese BPES family with four generations was enrolled in this study. Peripheral venous blood was collected from all affected patients, and genomic DNA was extracted from leukocytes. The whole coding sequence and nearby 5' untranslated region (UTR) and 3'UTR of the FOXL2 gene were amplified using polymerase chain reaction (PCR) with three sets of overlapping primers, followed by sequencing analyses. The sequencing results were analysed using SeqMan software. Based on the patients' clinical manifestations and analysis of the identified indel mutation, we found that the mutation disturbed interactions between FOXL2 and the StAR gene. Furthermore, through subcellular localisation and functional studies, we observed significant mislocalisation of the mutant protein; the mutant protein was found in the cytoplasm, while the wild-type protein was found in the nucleus. Loss of function was confirmed by transcriptional activity assays, quantitative real-time PCR, and electrophoretic mobility shift assays. Results: All affected patients presented with clinical features of BPES type I, including small palpebral fissures, ptosis, telecanthus, and epicanthus inversus with POF. A novel FOXL2 heterozygous indel mutation, c.19_95del, a 77-bp deletion that disrupts FOXL2 protein structure, was identified in all affected members of the family. In addition, this indel mutation significantly increased StAR mRNA expression by disrupting the ability of the FOXL2 protein to bind to the StAR promoter and act as a repressor of this gene. Conclusions: A novel FOXL2 indel mutation was identified in Chinese families with BPES. Our results expand the spectrum of known FOXL2 mutations and provide additional insight into the structure-function relationships of the FOXL2 protein. Furthermore, this novel mutation resulted in the dysfunction of FOXL2 as a transcription factor, blocking its ability to bind to the promoter region of the StAR gene, resulting in POF in the affected patient.
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影响因子：
3.5
作者：
Moumne, Lara;Dipietromaria, Aurelie;Veitia, Reiner A.
通讯作者：
Veitia, Reiner A.
影响因子：
3.7
作者：
Anttonen M;Pihlajoki M;Andersson N;Georges A;L'hôte D;Vattulainen S;Färkkilä A;Unkila-Kallio L;Veitia RA;Heikinheimo M
通讯作者：
Heikinheimo M
影响因子：
7.5
作者：
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通讯作者：
Gilks, C. Blake
影响因子：
4.8
作者：
Kuo, Fang-Ting;Bentsi-Barnes, Ikuko K.;Pisarska, Margareta D.
通讯作者：
Pisarska, Margareta D.
影响因子：
3.5
作者：
Uda, M;Ottolenghi, C;Pilia, G
通讯作者：
Pilia, G