Export from pericentriolar endocytic recycling compartment to cell surface depends on stable, detyrosinated (Glu) microtubules and kinesin
Export from pericentriolar endocytic recycling compartment to cell surface depends on stable, detyrosinated (Glu) microtubules and kinesin
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DOI：
10.1091/mbc.01-05-0224
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发表时间：
2002-01-01
影响因子：
3.3
通讯作者：
Maxfield, FR
中科院分区：
文献类型：
--
作者：
Lin, SX;Gundersen, GG;Maxfield, FR
一个显着部分的内化转铁蛋白（Tf）集中在内吞再循环室（ERC），这是附近的微管组织中心在许多细胞类型。然后转铁蛋白再回到细胞表面。控制ERC的定位、形态和功能的机制尚未完全了解。我们研究了转铁蛋白运输与微管（MT）的关系，特别是稳定的去酪氨酸Glu MT的子集。我们发现稳定的Glu MTs水平与ERC的分布之间存在一定的相关性;在中心粒附近Glu MTs水平较低的细胞中，ERC通常紧密聚集，而在整个细胞中Glu MTs水平较高的细胞中，ERC更为分散。紫杉醇处理后，随着Glu MTs水平的增加，中国仓鼠卵巢细胞中聚集的ERC变得分散。此外，在温度敏感的中国仓鼠卵巢细胞系（B104-5）中，当ERC在升高的温度下分散时，细胞具有更多的Glu MT。在高温下将纯化的抗Glu微管蛋白抗体微量注射到B104-5细胞中，诱导ERC重新分布为紧密的簇。微量注射抗Glu微管蛋白抗体可减缓转铁蛋白向细胞表面的再循环，而不影响转铁蛋白的内化或向ERC的传递。微量注射抗驱动蛋白抗体也能抑制转铁蛋白的再循环。这些结果表明，稳定的Glu MTs和驱动蛋白在ERC的组织中发挥作用，并促进囊泡从ERC移动到细胞表面。
A significant fraction of internalized transferrin (Tf) concentrates in the endocytic recycling compartment (ERC), which is near the microtubule-organizing center in many cell types. Tf then recycles back to the cell surface. The mechanisms controlling the localization, morphology, and function of the ERC are not fully understood. We examined the relationship of Tf trafficking with microtubules (MTs), specifically the subset of stable, detyrosinated Glu MTs. We found some correlation between the level of stable Glu MTs and the distribution of the ERC; in cells with low levels of Glu MTs concentrated near to the centriole, the ERC was often tightly clustered, whereas in cells with higher levels of Glu MTs throughout the cell, the ERC was more dispersed. The clustered ERC in Chinese hamster ovary cells became dispersed when the level of Glu MTs was increased with taxol treatment. Furthermore, in a temperature-sensitive Chinese hamster ovary cell line (B104-5), the cells had more Glu MTs when the ERC became dispersed at elevated temperature. Microinjecting purified anti-Glu tubulin antibody into B104-5 cells at elevated temperature induced the redistribution of the ERC to a tight cluster. Microinjection of anti-Glu tubulin antibody slowed recycling of Tf to the cell surface without affecting Tf internalization or delivery to the ERC. Similar inhibition of Tf recycling was caused by microinjecting anti-kinesin antibody. These results suggest that stable Glu MTs and kinesin play a role in the organization of the ERC and in facilitating movement of vesicles from the ERC to the cell surface.