Recycling endosomes can serve as intermediates during transport from the Golgi to the plasma membrane of MDCK cells.
Recycling endosomes can serve as intermediates during transport from the Golgi to the plasma membrane of MDCK cells.
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DOI：
10.1083/jcb.200408165
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发表时间：
2004-11-08
期刊：
影响因子：
--
通讯作者：
Mellman I
中科院分区：
文献类型：
--
作者：
Ang AL;Taguchi T;Francis S;Fölsch H;Murrells LJ;Pypaert M;Warren G;Mellman I
AP - 1B网格蛋白衔接蛋白复合物负责上皮细胞中许多基底外侧膜蛋白的极化运输。AP - 1B与参与AP - 1B依赖性运输的其他成分（胞吐复合体亚基和Rab8）一起定位于再循环内体（REs），这表明RE可能是高尔基体和质膜之间的中间体。尽管长期以来人们一直怀疑内体参与分泌途径，但我们现在使用四种独立的方法提供了直接证据，表明REs在MDCK细胞的基底外侧运输中起作用。通过视频显微镜、免疫电子显微镜和细胞分级分离发现，新合成的AP - 1B依赖性货物——水疱性口炎病毒糖蛋白G（VSV - G）在从反面高尔基体管网状结构离开后的几分钟内进入转铁蛋白阳性的REs。尽管是短暂的，但RE进入似乎是必不可少的，因为REs的酶失活会阻断VSV - G向细胞表面的运输。由于一种顶端靶向的VSV - G突变体表现相似，这些结果表明REs不仅作为中间体，而且作为内吞和分泌途径中极化分选的共同位点。
The AP-1B clathrin adaptor complex is responsible for the polarized transport of many basolateral membrane proteins in epithelial cells. Localization of AP-1B to recycling endosomes (REs) along with other components (exocyst subunits and Rab8) involved in AP-1B–dependent transport suggested that RE might be an intermediate between the Golgi and the plasma membrane. Although the involvement of endosomes in the secretory pathway has long been suspected, we now present direct evidence using four independent methods that REs play a role in basolateral transport in MDCK cells. Newly synthesized AP-1B–dependent cargo, vesicular stomatitis virus glycoprotein G (VSV-G), was found by video microscopy, immunoelectron microscopy, and cell fractionation to enter transferrin-positive REs within a few minutes after exit from the trans-Golgi network. Although transient, RE entry appears essential because enzymatic inactivation of REs blocked VSV-G delivery to the cell surface. Because an apically targeted VSV-G mutant behaved similarly, these results suggest that REs not only serve as an intermediate but also as a common site for polarized sorting on the endocytic and secretory pathways.
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