Skirball Institute of Biomolecular Medicine, NYU Grossman School of Medicine, New York, United States.
Department of Cell Biology, NYU Grossman School of Medicine, New York, United States.
Elife. 2021 Mar 9;10:e65169. doi: 10.7554/eLife.65169.
Lumen extension in intracellular tubes can occur when vesicles fuse with an invading apical membrane. Within the excretory cell, which forms an intracellular tube, the exocyst vesicle-tethering complex is enriched at the lumenal membrane and is required for its outgrowth, suggesting that exocyst-targeted vesicles extend the lumen. Here, we identify a pathway that promotes intracellular tube extension by enriching the exocyst at the lumenal membrane. We show that PAR-6 and PKC-3/aPKC concentrate at the lumenal membrane and promote lumen extension. Using acute protein depletion, we find that PAR-6 is required for exocyst membrane recruitment, whereas PAR-3, which can recruit the exocyst in mammals, appears dispensable for exocyst localization and lumen extension. Finally, we show that CDC-42 and RhoGEF EXC-5/FGD regulate lumen extension by recruiting PAR-6 and PKC-3 to the lumenal membrane. Our findings reveal a pathway that connects CDC-42, PAR proteins, and the exocyst to extend intracellular tubes.
当囊泡与入侵的顶膜融合时,细胞内管的腔延伸会发生。在形成细胞内管的分泌细胞中,外泌体囊泡连接复合物在腔膜处富集,并需要其生长,这表明外泌体靶向囊泡延伸了腔。在这里,我们确定了一条通过在腔膜处富集外泌体来促进细胞内管延伸的途径。我们表明 PAR-6 和 PKC-3/aPKC 集中在腔膜上,并促进腔延伸。通过急性蛋白耗竭,我们发现 PAR-6 是外泌体膜募集所必需的,而在哺乳动物中可以募集外泌体的 PAR-3 对于外泌体定位和腔延伸似乎是可有可无的。最后,我们表明 CDC-42 和 RhoGEF EXC-5/FGD 通过将 PAR-6 和 PKC-3 募集到腔膜上来调节腔延伸。我们的发现揭示了一条连接 CDC-42、PAR 蛋白和外泌体以延伸细胞内管的途径。