Young Alejandra, Dandekar Uma, Pan Calvin, Sader Avery, Zheng Jie J, Lewis Richard A, Farber Debora B
Stein Eye Institute and Department of Ophthalmology, David Geffen School of Medicine, UCLA, Los Angeles, CA, United States of America.
Molecular Biology Institute, UCLA, Los Angeles, CA, United States of America.
PLoS One. 2016 Sep 8;11(9):e0162273. doi: 10.1371/journal.pone.0162273. eCollection 2016.
Ocular albinism type 1 (OA), caused by mutations in the OA1 gene, encodes a G-protein coupled receptor, OA1, localized in melanosomal membranes of the retinal pigment epithelium (RPE). This disorder is characterized by both RPE macro-melanosomes and abnormal decussation of ganglion cell axons at the brain's optic chiasm. We demonstrated previously that Oa1 specifically activates Gαi3, which also signals in the Oa1 transduction pathway that regulates melanosomal biogenesis. In this study, we screened the human Gαi3 gene, GNAI3, in DNA samples from 26 patients who had all clinical characteristics of OA but in whom a specific mutation in the OA1 gene had not been found, and in 6 normal control individuals. Using the Agilent HaloPlex Target Enrichment System and next-generation sequencing (NGS) on the Illumina MiSeq platform, we identified 518 variants after rigorous filtering. Many of these variants were corroborated by Sanger sequencing. Overall, 98.8% coverage of the GNAI3 gene was obtained by the HaloPlex amplicons. Of all variants, 6 non-synonymous and 3 synonymous were in exons, 41 in a non-coding exon embedded in the 3' untranslated region (UTR), 6 in the 5' UTR, and 462 in introns. These variants included novel SNVs, insertions, deletions, and a frameshift mutation. All were found in at least one patient but none in control samples. Using computational methods, we modeled the GNAI3 protein and its non-synonymous exonic mutations and determined that several of these may be the cause of disease in the patients studied. Thus, we have identified GNAI3 as a second gene possibly responsible for X-linked ocular albinism.
1型眼白化病(OA)由OA1基因突变引起,该基因编码一种G蛋白偶联受体OA1,定位于视网膜色素上皮(RPE)的黑素小体膜上。这种疾病的特征是RPE大黑素小体以及神经节细胞轴突在脑视交叉处的异常交叉。我们之前证明Oa1特异性激活Gαi3,Gαi3也在调节黑素小体生物发生的Oa1转导途径中发挥信号作用。在本研究中,我们对26例具有OA所有临床特征但未发现OA1基因特定突变的患者以及6例正常对照个体的DNA样本进行了人类Gαi3基因(GNAI3)筛查。使用安捷伦HaloPlex目标富集系统和Illumina MiSeq平台上的新一代测序(NGS),经过严格筛选后我们鉴定出518个变体。其中许多变体通过桑格测序得到了证实。总体而言,HaloPlex扩增子对GNAI3基因的覆盖度达到了98.8%。在所有变体中,有6个非同义变体和3个同义变体在外显子中,41个在3'非翻译区(UTR)嵌入的非编码外显子中,6个在5'UTR中,462个在内含子中。这些变体包括新的单核苷酸变异(SNV)、插入、缺失和一个移码突变。所有这些变体至少在一名患者中被发现,而在对照样本中均未发现。我们使用计算方法对GNAI3蛋白及其非同义外显子突变进行建模,并确定其中一些可能是所研究患者疾病的病因。因此,我们已将GNAI3鉴定为可能导致X连锁眼白化病的第二个基因。