Shared as well as distinct roles of EHD proteins revealed by biochemical and functional comparisons in mammalian cells and C. elegans.
Shared as well as distinct roles of EHD proteins revealed by biochemical and functional comparisons in mammalian cells and C. elegans.
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哺乳动物细胞和秀丽隐杆线虫中的生化和功能比较揭示了EHD蛋白的共享和不同作用。
DOI：
10.1186/1471-2121-8-3
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发表时间：
2007-01-18
期刊：
影响因子：
--
通讯作者：
Band, Hamid
中科院分区：
文献类型：
--
作者：
George, Manju;Ying, GuoGuang;Rainey, Mark A.;Solomon, Aharon;Parikh, Pankit T.;Gao, Qingshen;Band, Vimla;Band, Hamid
四种高度同源的人EHD蛋白（EHD1-4）形成了Eps15同源结构域蛋白家族的不同亚家族，并被认为调节内吞循环。这个家族的某些成员已经在不同的细胞背景下进行了研究；然而，缺乏对所有四种蛋白质的同步分析阻碍了对它们的冗余与不同功能的欣赏。在这里，我们分析了哺乳动物细胞中的四种EHD蛋白，并使用缺乏EHD同源物RME-1的秀丽隐杆线虫突变体进行了跨物种互补实验。研究人员发现，所有人类EHD蛋白都能拯救线虫rme-1突变体的空泡化肠道表型，并在一组哺乳动物细胞系和组织中同时表达，并与Rab11（一种循环核内体标记物）和同源或异质寡聚和共定位。小干扰RNA （siRNA）敲除EHD1、2和4，以及EH结构域显性阴性缺失突变体的表达表明，EHD1和3（以及较小程度上的EHD4）的缺失，而不是EHD2功能的缺失，会延迟转铁蛋白从内噬循环室的退出。EH结构域缺失突变的EHD1和3，而不是2和4，诱导了显著的共转染Rab11的核周围聚集。敲除分析表明，EHD1和2调节货物从回收内体的出口，而EHD4，类似于EHD3的报道（Naslavsky et al. (2006) Mol. Biol）。细胞17,163)，调节从早期核内体到再循环核内体的运输。总之，我们的研究表明，同时表达的人EHD蛋白在内吞循环途径中发挥着共享和离散的功能，为未来研究识别和表征所涉及的分子途径奠定了基础。
The four highly homologous human EHD proteins (EHD1-4) form a distinct subfamily of the Eps15 homology domain-containing protein family and are thought to regulate endocytic recycling. Certain members of this family have been studied in different cellular contexts; however, a lack of concurrent analyses of all four proteins has impeded an appreciation of their redundant versus distinct functions. Here, we analyzed the four EHD proteins both in mammalian cells and in a cross-species complementation assay using a C. elegans mutant lacking the EHD ortholog RME-1. We show that all human EHD proteins rescue the vacuolated intestinal phenotype of C. elegans rme-1 mutant, are simultaneously expressed in a panel of mammalian cell lines and tissues tested, and variably homo- and hetero-oligomerize and colocalize with each other and Rab11, a recycling endosome marker. Small interfering RNA (siRNA) knock-down of EHD1, 2 and 4, and expression of dominant-negative EH domain deletion mutants showed that loss of EHD1 and 3 (and to a lesser extent EHD4) but not EHD2 function retarded transferrin exit from the endocytic recycling compartment. EH domain deletion mutants of EHD1 and 3 but not 2 or 4, induced a striking perinuclear clustering of co-transfected Rab11. Knock-down analyses indicated that EHD1 and 2 regulate the exit of cargo from the recycling endosome while EHD4, similar to that reported for EHD3 (Naslavsky et al. (2006) Mol. Biol. Cell 17, 163), regulates transport from the early endosome to the recycling endosome. Altogether, our studies suggest that concurrently expressed human EHD proteins perform shared as well as discrete functions in the endocytic recycling pathway and lay a foundation for future studies to identify and characterize the molecular pathways involved.
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影响因子：
3.3
作者：
Lin, SX;Gundersen, GG;Maxfield, FR
通讯作者：
Maxfield, FR
DOI：
10.1016/s0006-291x(02)00282-6
发表时间：
2002-05-03
影响因子：
3.1
作者：
Iwahashi, J;Kawasaki, I;Toyoda, T
通讯作者：
Toyoda, T
影响因子：
4.8
作者：
Lee, DW;Zhao, XH;Greene, LE
通讯作者：
Greene, LE
影响因子：
4
作者：
Grimberg, E;Peng, Z;Sagi-Eisenberg, R
通讯作者：
Sagi-Eisenberg, R
影响因子：
9.2
作者：
Mayer, BJ
通讯作者：
Mayer, BJ