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Identification of Genes Required for Apical Protein Trafficking in Drosophila Photoreceptor Cells

机译:鉴定果蝇感光细胞顶蛋白运输所需的基因。

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摘要

photoreceptor cells are highly polarized epithelial cells. Their apical membrane is further subdivided into the stalk membrane and the light-sensing rhabdomere. The photo-pigment Rhodopsin1 (Rh1) localizes to the rhabdomere, whereas the apical determinant Crumbs (Crb) is enriched at the stalk membrane. The proteoglycan Eyes shut (Eys) is secreted through the apical membrane into an inter-rhabdomeral space. Rh1, Crb, and Eys are essential for the development of photoreceptor cells, normal vision, and photoreceptor cell survival. Human orthologs of all three proteins have been linked to retinal degenerative diseases. Here, we describe an RNAi-based screen examining the importance of 237 trafficking-related genes in apical trafficking of Eys, Rh1, and Crb. We found 28 genes that have an effect on the localization and/or levels of these apical proteins and analyzed several factors in more detail. We show that the Arf GEF protein Sec71 is required for biosynthetic traffic of both apical and basolateral proteins, that the exocyst complex and the microtubule-based motor proteins dynein and kinesin promote the secretion of Eys and Rh1, and that Syntaxin 7/Avalanche controls the endocytosis of Rh1, Eys, and Crb.
机译:感光细胞是高度极化的上皮细胞。它们的顶膜可进一步细分为茎膜和光感横纹肌。光色素Rhodopsin1(Rh1)定位在横纹肌上,而顶端决定簇Crumbs(Crb)富集在茎膜上。蛋白糖闭眼(Eys)通过顶膜分泌到横纹肌间隙。 Rh1,Crb和Eys对于感光细胞的发育,正常视力和感光细胞的存活至关重要。所有这三种蛋白质的人类直系同源物都与视网膜变性疾病有关。在这里,我们描述了一个基于RNAi的屏幕,该屏幕检查了237种与运输有关的基因在Eys,Rh1和Crb的顶端运输中的重要性。我们发现了28个对这些顶端蛋白质的定位和/或水平有影响的基因,并更详细地分析了一些因素。我们显示Arf GEF蛋白Sec71是顶端和基底外侧蛋白的生物合成运输所必需的,囊外复合物和基于微管的动力蛋白dynein和kinesin促进Eys和Rh1的分泌,而Syntaxin 7 / Avalanche控制着Rh1,Eys和Crb的胞吞作用。

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