Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, Doisy Research Center, St. Louis, MO 63104, USA.
Traffic. 2011 Dec;12(12):1821-38. doi: 10.1111/j.1600-0854.2011.01285.x. Epub 2011 Oct 13.
Golgi-localized, γ-ear-containing, ADP ribosylation factor-binding (GGA) proteins are monomeric adaptors implicated in clathrin-mediated vesicular transport between the trans Golgi network and endosomes, characterized mainly from cell culture analysis of lysosomal sorting. To provide the first demonstration of GGA's role in vivo, we used Drosophila which has a single GGA and a single lysosomal sorting receptor, lysosomal enzyme receptor protein (LERP). Using RNAi knockdowns, we show that the Drosophila GGA is required for lysosomal sorting. We further identified authentic components of the Drosophila lysosomal sorting system--the sorting receptor LERP, the sorting adaptor GGA and the lysosomal cargo cathepsins B1, D and L--to show that GGA depletion results in lysosomal dysfunction. Abnormal lysosomal morphology, missorting of lysosomal cathepsins and impaired lysosomal proteolysis show disturbed LERP trafficking after GGA depletion. GGA is highly expressed in the mushroom bodies and the pigment cells of the retina, and increasing or decreasing the levels of GGA in the eyes leads to retinal defects. Reduced GGA levels also enhance an eye defect caused by overexpression of the autophagy-associated protein Blue cheese (Bchs), implicating GGA in autophagic processes. This shows that Drosophila provides an excellent whole-animal model to gain new insights into the function of GGA in the physiological environment of a multicellular organism.
高尔基定位、γ 耳含有、ADP 核糖基化因子结合(GGA)蛋白是单体衔接蛋白,参与网格蛋白包被小泡运输从反高尔基网络到内体之间的运输,主要从溶酶体分选的细胞培养分析中得到鉴定。为了首次在体内证明 GGA 的作用,我们使用了果蝇,它只有一个 GGA 和一个溶酶体分选受体,溶酶体酶受体蛋白(LERP)。通过 RNAi 敲低,我们表明果蝇 GGA 是溶酶体分选所必需的。我们进一步鉴定了果蝇溶酶体分选系统的真实成分——分选受体 LERP、分选衔接蛋白 GGA 和溶酶体货物组织蛋白酶 B1、D 和 L——表明 GGA 耗竭导致溶酶体功能障碍。异常的溶酶体形态、溶酶体组织蛋白酶的错误分选以及溶酶体蛋白水解的受损表明 GGA 耗竭后 LERP 转运受到干扰。GGA 在蘑菇体和视网膜的色素细胞中高度表达,增加或减少眼睛中的 GGA 水平会导致视网膜缺陷。降低 GGA 水平还会增强自噬相关蛋白 Blue cheese(Bchs)过表达引起的眼部缺陷,表明 GGA 参与自噬过程。这表明果蝇提供了一个极好的整体动物模型,可深入了解 GGA 在多细胞生物体生理环境中的功能。