The centrosome regulates the Rab11- dependent recycling endosome pathway at appendages of the mother centriole.

The centrosome regulates the Rab11- dependent recycling endosome pathway at appendages of the mother centriole.
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DOI:
10.1016/j.cub.2012.08.022
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发表时间:
2012-10-23
期刊:
影响因子:
9.2
通讯作者:
Doxsey, Stephen
Doxsey, Stephen
中科院分区:
生物学1区
文献类型:
--
作者:
Hehnly, Heidi;Chen, Chun-Ting;Powers, Christine M.;Liu, Hui-Lin;Doxsey, Stephen

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再循环核内体通过微管依赖的运输定位于核内体周围区域。我们之前的研究表明,再生核内体成分Rab11-GTPase的效应物Sec15与母体中心粒附属蛋白中心粒蛋白相互作用,表明核内体和中心体之间存在相互作用。在这里,我们显示了再循环核内体与母体(老)中心粒的附属物相结合。研究表明,中心粒母体附属蛋白,中心粒蛋白和中心粒蛋白/ODF2,调节内核体成分Rab11、Rab11 gtp激活蛋白Evi5和母中心粒囊的结合。在分离的无膜中心体上重建内核体蛋白复合物的体外方法表明,纯化的GTP-Rab11在没有膜的情况下与母中心粒附着物结合,而不是GDP-Rab11。此外,中心粒蛋白耗竭使中心体Rab11 GAP、Evi5发生位移,并增加母体-中心粒相关的Rab11;Evi5的缺失也增加了中心体Rab11。这表明中心蛋白将Evi5定位到中心粒附属物上,以关闭中心体Rab11的活性。最后,中心蛋白的缺失破坏了再循环核内体的组织和功能,这表明母中心粒蛋白在调控Rab11在母中心粒的定位和活性方面发挥了作用。
The recycling endosome localizes to a pericentrosomal region via microtubule-dependent transport. We previously showed that Sec15, an effector of the recycling endosome component, Rab11-GTPase, interacts with the mother centriole appendage protein, centriolin, suggesting an interaction between endosomes and centrosomes. Here we show that the recycling endosome associates with the appendages of the mother (older) centriole. We show that the mother centriole appendage proteins, centriolin and cenexin/ODF2, regulate association of the endosome components Rab11, the Rab11 GTP-activating protein Evi5, and the exocyst at the mother centriole. Development of an in vitro method for reconstituting endosome protein complexes onto isolated membrane-free centrosomes demonstrates that purified GTP-Rab11 but not GDP-Rab11 binds to mother centriole appendages in the absence of membranes. Moreover, centriolin depletion displaces the centrosomal Rab11 GAP, Evi5, and increases mother-centriole-associated Rab11; depletion of Evi5 also increases centrosomal Rab11. This indicates that centriolin localizes Evi5 to centriolar appendages to turn off centrosomal Rab11 activity. Finally, centriolin depletion disrupts recycling endosome organization and function suggesting a role for mother centriole proteins in the regulation of Rab11 localization and activity at the mother centriole.
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