MOLECULAR BIOLOGY OF OUTER RETINA-SPECIFIC PROTEINS

视网膜色素上皮（RPE）细胞和感光细胞是 功能和发展紧密结合。 第一章 RPE与某些视网膜疾病有关。 然而，RPE很差， 在分子水平上理解。 我们正在表征RPE 65，a 发育调节的、保守的65 kD RPE特异性微粒体 膜相关蛋白 我们克隆了RPE 65的cDNA，发现 它编码了一种新的蛋白质。 这种蛋白质没有任何 预测的跨膜片段，但它具有很强的亲和力， 磷脂，这可能与其功能有关。 cdna序列 用于过表达RPE 65蛋白进行功能研究。 的 还将研究该蛋白在诱导葡萄膜炎中的潜在作用 使用重组蛋白。 在培养的RPE细胞中RPE 65 mRNA的翻译缺乏， 作为一种可能的转录后调节机制进行研究 这可能与RPE-视网膜关系以及RPE 移植研究。 我们分离出了一个全长基因组克隆 RPE65 它的长度至少为22 kb。 我们使用了cDNA， 将人RPE 65基因定位于染色体1p31的基因组序列 而小鼠的同源基因则位于远端3号染色体。 这些并不符合 与目前发现的任何眼部疾病基因都有关联 然而，RPE65仍然存在， RPE相关疾病的候选基因。

Retinal pigment epithelium (RPE) cells and photoreceptor cells are functionally and developmentally closely integrated. Derangements of the RPE are involved in certain retinal diseases. However, the RPE is poorly understood at the molecular level. We are characterizing RPE65, a developmentally regulated, conserved 65-kD RPE-specific microsomal membrane-associated protein. We have cloned the cDNA for RPE65 and found that it encodes a novel protein. This protein does not have any predicted transmembrane segments, yet it has a strong affinity for phospholipids, which may be related to its function. The cDNA sequence is being used to overexpress RPE65 protein for functional studies. The potential role of the protein in inducing uveitis also will be studied using recombinant protein. The lack of translation of RPE65 mRNA in cultured RPE cells is being investigated as a possible mechanism of posttranscriptional regulation that may have a bearing on the RPE-retina relationship s well as on RPE transplantation studies. We have isolated a full-length genomic clone for RPE65. It is at least 22 kb in length. We have used the cDNA and genomic sequences to localize the human gene for RPE65 to chromosome 1p31 and the mouse homologue to distal chromosome 3. These do not correspond to any ocular disease gene localized so far. Nonetheless, RPE65 remains a candidate gene for RPE-involved disease.