CeVPS-27 is an endosomal protein required for the molting and the endocytic trafficking of the low-density lipoprotein receptor-related protein 1 in Caenorhabditis elegans
CeVPS-27 is an endosomal protein required for the molting and the endocytic trafficking of the low-density lipoprotein receptor-related protein 1 in Caenorhabditis elegans
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DOI：
10.1111/j.1600-0854.2005.00309.x
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发表时间：
2005-08-01
期刊：
影响因子：
4.5
通讯作者：
Legouis, R
中科院分区：
文献类型：
--
作者：
Roudier, N;Lefebvre, C;Legouis, R
E 类液泡蛋白分选 (Vps) 蛋白首次在酵母中被描述为参与受体介导的内吞作用和多泡体形成。通过 RNA 干扰使线虫秀丽隐杆线虫 E 类 VPS 基因失活，揭示了异质表型。我们进一步表征了必需基因 Cevps-27（人肝细胞生长因子调节的酪氨酸激酶底物的直系同源物）在秀丽隐杆线虫发育过程中的作用。使用绿色荧光蛋白融合构建体和抗体染色显示 Cevps-27 定位于内体膜。它广泛表达但在上皮细胞中富集。电子显微镜和自噬标记 LGG-1 分析显示，Cevps-27 突变体呈现扩大的内体结构和自噬囊泡的积累。 Cevps-27 动物在 L2-L3 蜕皮时被捕，无法降解其旧角质层。当 Cevps-27 蠕虫在次优浓度的胆固醇下生长时，这种蜕皮表型更加严重。此外，在 Cevps-27 突变体中还观察到低密度脂蛋白受体相关蛋白 1 (LRP-1) 的内吞运输缺陷。这些结果表明，CeVPS-27 是线虫内体和自噬途径所必需的，并且在通过 LRP-1 内化和胆固醇运输控制蜕皮中发挥着至关重要的作用。
Class E vacuolar protein-sorting (Vps) proteins were first described in yeast as being involved in receptor-mediated endocytosis and multivesicular body formation. Inactivation by RNA interference of the class E VPS genes of the nematode Caenorhabditis elegans revealed heterogeneous phenotypes. We have further characterized the role of the essential gene Cevps-27, ortholog of human hepatocyte growth factor-regulated tyrosine kinase substrate, during the development of C. elegans. Use of green fluorescent protein fusion constructs and antibody staining revealed that Cevps-27 localizes to endosomal membranes. It is widely expressed but enriched in epithelial cells. Cevps-27 mutants presented enlarged endosomal structures and an accumulation of autophagic vesicles as revealed by electron microscopy and the analysis of the autophagic marker LGG-1. Cevps-27 animals arrested at L2-L3 molt with an inability to degrade their old cuticle. This molting phenotype was more severe when Cevps-27 worms were grown on suboptimal concentrations of cholesterol. Furthermore, defective endocytic trafficking of the low-density lipoprotein receptor-related protein 1 (LRP-1) was also observed in Cevps-27 mutants. These results indicate that CeVPS-27 is required for endosomal and autophagic pathways in C. elegans and plays a crucial role in the control of molting through LRP-1 internalization and cholesterol traffic.