Intra‐ and interspecies comparison of EYS transcripts highlights its characteristics in the eye
Intra‐ and interspecies comparison of EYS transcripts highlights its characteristics in the eye
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DOI：
10.1096/fj.201900056rr
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发表时间：
2019-05
期刊：
影响因子：
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通讯作者：
Shimpei Takita;Kiyoko Miyamoto-Matsui;Y. Seko
中科院分区：
文献类型：
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作者：
Shimpei Takita;Kiyoko Miyamoto-Matsui;Y. Seko
闭眼同源基因 (EYS) 的遗传性突变会导致色素性视网膜炎。尽管斑马鱼眼的敲除具有致病性，但脊椎动物光感受器中 EYS 的分子功能仍知之甚少。在这里，我们证明 EYS 的 5' 部分在脊椎动物中具有眼睛特异性。我们之前确定，具有未知转录起始位点的 EYS 3' 片段在人真皮成纤维细胞 (HDF) 中表达。为了深入了解 EYS 在脊椎动物光感受器中的分子功能，我们使用 cDNA 末端快速扩增、基因表达帽分析、RNA 测序和 RT-PCR 广泛分析了人成纤维细胞系 HDF-adult (HDF-a)、Y79 视网膜母细胞瘤细胞系和斑马鱼眼睛中的 EYS (eys) 表达。在 HDF-a 细胞中，我们发现了一种新的转录变体 (tv)，tv5，从外显子 37 转录。在 Y79 细胞和斑马鱼眼睛中，EYS (eys) 主要从外显子 1 或 2 转录，而在 HDF-a 细胞中，它仅从外显子 37 转录。在斑马鱼眼睛中，存在引入终止密码子的剪接变体，导致 RNA 3' 部分完全丢失。这些比较方法表明，EYS (eys) mRNA 的 5' 部分似乎具有光感受器特异性，并且推导的眼睛中 EYS 蛋白的组成在脊椎动物中非常保守。—Takita, S.、Miyamoto-Matsui, K.、Seko, Y。EYS 转录本的种内和种间比较突出了其在眼睛中的特征。 FASEB J. 33, 9422–9433 (2019)。 www.fasebj.org
Inherited mutations in the eyes shut homolog (EYS) gene cause retinitis pigmentosa. Although knock out of eys in zebrafish is pathogenic, the molecular function of EYS in vertebrate photoreceptors is poorly understood. Here, we show that the 5′ portion of EYS is eye‐specific across vertebrates. We previously determined that a 3′ fragment of EYS with an unknown transcription start site is expressed in human dermal fibroblasts (HDF). To obtain insights into the molecular function of EYS in vertebrate photoreceptors, we extensively analyzed EYS (eys) expression in the human fibroblast cell line HDF‐adult (HDF‐a), the Y79 retinoblastoma cell line, and in zebrafish eyes using rapid amplification of cDNA end, cap analysis of gene expression, RNA sequencing, and RT‐PCR. In HDF‐a cells, we identified a novel transcript variant (tv), tv5, transcribed from exon 37. In Y79 cells and zebrafish eyes, EYS (eys) was predominantly transcribed from exon 1 or 2, whereas it was transcribed exclusively from exon 37 in HDF‐a cells. In the zebrafish eye, there were splice variants that introduced stop codons, resulting in complete loss of the 3′ portion of the RNA. These comparative approaches indicate that the 5′ portion of the EYS (eys) mRNA appears to be photoreceptor‐specific and that the compositions of the deduced EYS proteins in the eye are well‐conserved across vertebrates.—Takita, S., Miyamoto‐Matsui, K., Seko, Y. Intra‐ and interspecies comparison of EYS transcripts highlights its characteristics in the eye. FASEB J. 33, 9422–9433 (2019). www.fasebj.org