Interaction between optineurin and the bZIP transcription factor NRL
Interaction between optineurin and the bZIP transcription factor NRL
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DOI：
10.1002/cbin.10174
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发表时间：
2009-04
影响因子：
3.9
通讯作者：
Chunxia Wang;Katsuhiro Hosono;M. Ohtsubo;Kentaro Ohishi;Jie Gao;Nobuo Nakanishi;Akiko Hikoya;Miho Sato;Y. Hotta;S. Minoshima
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文献类型：
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作者：
Chunxia Wang;Katsuhiro Hosono;M. Ohtsubo;Kentaro Ohishi;Jie Gao;Nobuo Nakanishi;Akiko Hikoya;Miho Sato;Y. Hotta;S. Minoshima
虽然编码视神经磷酸酶（OPTN）的基因是青光眼和肌萎缩侧索硬化症的致病基因，但它在所有身体组织中普遍表达，包括视网膜。为了研究OPTN在视网膜神经节细胞以及整个视网膜中的功能，我们先前分离了OPTN相互作用蛋白，并鉴定了编码bZIP转录因子神经视网膜亮氨酸拉链（NRL）的基因，该基因是视网膜色素变性的致病基因。在此，我们研究了在HeLaS 3细胞中OPTN和NRL蛋白之间的结合。HA标记的NRL和FLAG标记的OPTN在HeLaS 3细胞中的共表达，随后用抗标签抗体进行免疫沉淀和蛋白质印迹，证明了这些蛋白质在HeLaS 3细胞中的结合，这通过邻位连接试验得到证实。NRL是第一个表现出眼特异性表达的OPTN结合蛋白。一系列部分缺失OPTN质粒证明OPTN的尾区（423-577个氨基酸[aa]）是与NRL结合所必需的。免疫组化结果显示，Optn（大鼠OPTN同源物）在大鼠感光细胞中表达，并定位于感光细胞的细胞质中。这是光感受器细胞中Optn表达的新证明。在我们的实验条件下，在感光细胞核中未检测到OPTN。进一步的分析是必要的，以阐明OPTN的功能及其可能与感光细胞中的NRL结合的意义。
Although the gene encoding optineurin (OPTN) is a causative gene for glaucoma and amyotrophic lateral sclerosis, it is ubiquitously expressed in all body tissues, including the retina. To study the function of OPTN in retinal ganglion cells as well as the whole retina, we previously isolated OPTN‐interacting proteins and identified the gene encoding the bZIP transcription factor neural retina leucine zipper (NRL), which is a causative gene for retinitis pigmentosa. Herein, we investigated the binding between OPTN and NRL proteins in HeLaS3 cells. Co‐expression of HA‐tagged NRL and FLAG‐tagged OPTN in HeLaS3 cells followed by immunoprecipitation and Western blotting with anti‐tag antibodies demonstrated the binding of these proteins in HeLaS3 cells, which was confirmed by proximity ligation assay. NRL is the first OPTN‐binding protein to show eye‐specific expression. A series of partial‐deletion OPTN plasmids demonstrated that the tail region (423–577 amino acids [aa]) of OPTN was necessary for binding with NRL. Immunostaining showed that Optn (rat homologue of OPTN) was expressed in rat photoreceptors and localised in the cytoplasm of photoreceptor cells. This is a novel demonstration of Optn expression in photoreceptor cells. OPTN was not detected in photoreceptor nuclei under our experimental conditions. Further analyses are necessary to elucidate the function of OPTN and the significance of its possible binding with NRL in photoreceptor cells.