A novel mutation of p.F32I in GJA8 in human dominant congenital cataracts
A novel mutation of p.F32I in GJA8 in human dominant congenital cataracts
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人类显性先天性白内障 GJA8 中 p.F32I 的新突变
DOI：
10.18240/ijo.2016.11.05
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发表时间：
2016-11-18
影响因子：
1.4
通讯作者：
Zhu, Yi-Hua
中科院分区：
文献类型：
--
作者：
Dang, Feng-Tao;Yang, Fa-Yu;Zhu, Yi-Hua
目的：目的：鉴定一个三代常染色体显性遗传性先天性全白内障家系的致病基因突变，并分析该突变的分子遗传学后果。通过聚合酶链反应（PCR）筛选与人类总白内障相关的两个已报道基因（GRYAA和GJA8）的突变，并对PCR产物进行直接测序。分析了鉴定的突变的分子后果。产生携带编码连接蛋白50（Cx50）的Gja 8的野生型和突变型小鼠ORF的质粒，并在293细胞中异位表达。共聚焦显微镜检测Cx50蛋白表达和细胞定位。结果：对患病和未患病的家族成员进行临床和眼科检查。通过对与人类全白内障相关的两个已报道基因（GRYAA和GJA8）进行PCR并对PCR产物进行直接测序，在招募的家族成员中筛选突变。分析了鉴定的突变的分子后果。产生携带编码Cx50的Gja 8的野生型和突变型小鼠ORF的质粒，并在293细胞中异位表达。通过共聚焦显微镜对重组Cx50的重组蛋白表达和细胞定位进行了评估。结论：本研究确定了GJA8中的一种新的白内障突变，这为现有的Cx50突变与白内障的谱增加了一种新的突变。GJA8中p.F32I突变的分子后果排除了突变型Cx50蛋白的不稳定性和错误定位。
AIM: To identify a causative mutation in a three generation family with autosomal dominant congenital total cataract and dissect the molecular consequence of the identified mutation.METHODS: Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by polymerase chain reaction (PCR) of the two reported genes (GRYAA and GJA8) which were linked to human total cataracts and direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected. The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for connexin 50 (Cx50), were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy.RESULTS: Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by PCR of the two reported genes (GRYAA and GJA8) which were linked to human total cataracts and direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected. The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for Cx50, were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy.CONCLUSION: This study has identified a novel cataract mutation in GJA8, which adds a novel mutation to the existing spectrum of Cx50 mutations with cataract. The molecular consequences of p.F32I mutation in GJA8 exclude instability and the mislocalization of mutant Cx50 protein.