EHBP-1 functions with RAB-10 during endocytic recycling in Caenorhabditis elegans.
EHBP-1 functions with RAB-10 during endocytic recycling in Caenorhabditis elegans.
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DOI：
10.1091/mbc.e10-02-0149
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发表时间：
2010-08-15
影响因子：
3.3
通讯作者：
Grant BD
中科院分区：
文献类型：
--
作者：
Shi A;Chen CC;Banerjee R;Glodowski D;Audhya A;Rongo C;Grant BD
秀丽隐杆线虫RAB-10在极化细胞中的内吞再循环中起作用，调节肠上皮中的基底外侧货物运输和中间神经元中的突触后货物运输。在这里，我们显示RAB-10的结合EHBP-1，CH-结构域蛋白，并证明EHBP-1的RAB-10调节的运输在这两种组织中的要求。秀丽隐杆线虫RAB-10在极化细胞中的内吞再循环中起作用，调节肠上皮中的基底外侧货物运输和中间神经元中的突触后货物运输。哺乳动物Rab 10在MDCK细胞中也有类似的作用，这表明在后生动物中有一种保守的机制调节这些相关的通路。在酵母双杂交筛选RAB-10的结合伙伴，我们确定了EHBP-1，钙调蛋白同源结构域（CH）蛋白，其哺乳动物同系物Ehbp 1先前被证明在脂肪细胞内吞转运GLUT 4的功能。在体内，我们发现EHBP-1-GFP与RFP-RAB-10共定位在肠和中间神经元的内体结构上，并且ehbp-1功能丧失突变体与rab-10突变体共享特定的内体形态和货物定位缺陷。我们还表明，EHBP-1的损失破坏了膜蛋白运输到质膜的非极化生殖细胞，一个缺陷，可以表型复制的编码RAB-10和它的最接近的parparaplation RAB-8。这些结果表明，RAB-10和EHBP-1在许多细胞类型中一起发挥作用，并表明Rab家族成员在极化细胞与非极化细胞中的冗余水平存在差异。
Caenorhabditis elegans RAB-10 functions in endocytic recycling in polarized cells, regulating basolateral cargo transport in the intestinal epithelia and postsynaptic cargo transport in interneurons. Here we show binding of RAB-10 to EHBP-1, a CH-domain protein, and demonstrate a requirement for EHBP-1 in RAB-10–regulated transport in both of these tissues. Caenorhabditis elegans RAB-10 functions in endocytic recycling in polarized cells, regulating basolateral cargo transport in the intestinal epithelia and postsynaptic cargo transport in interneurons. A similar role was found for mammalian Rab10 in MDCK cells, suggesting that a conserved mechanism regulates these related pathways in metazoans. In a yeast two-hybrid screen for binding partners of RAB-10 we identified EHBP-1, a calponin homology domain (CH) protein, whose mammalian homolog Ehbp1 was previously shown to function during endocytic transport of GLUT4 in adipocytes. In vivo we find that EHBP-1-GFP colocalizes with RFP-RAB-10 on endosomal structures of the intestine and interneurons and that ehbp-1 loss-of-function mutants share with rab-10 mutants specific endosome morphology and cargo localization defects. We also show that loss of EHBP-1 disrupts transport of membrane proteins to the plasma membrane of the nonpolarized germline cells, a defect that can be phenocopied by codepletion of RAB-10 and its closest paralog RAB-8. These results indicate that RAB-10 and EHBP-1 function together in many cell types and suggests that there are differences in the level of redundancy among Rab family members in polarized versus nonpolarized cells.