Rabankyrin-5 interacts with EHD1 and Vps26 to regulate endocytic trafficking and retromer function.
Rabankyrin-5 interacts with EHD1 and Vps26 to regulate endocytic trafficking and retromer function.
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DOI：
10.1111/j.1600-0854.2012.01334.x
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发表时间：
2012-05
期刊：
影响因子：
--
通讯作者：
Caplan S
中科院分区：
文献类型：
--
作者：
Zhang J;Reiling C;Reinecke JB;Prislan I;Marky LA;Sorgen PL;Naslavsky N;Caplan S
Rabankyrin-5(Rank-5)被认为是小GTP酶Rab5的效应因子，在巨噬细胞增多中起重要作用。我们现在已经确定Rank-5是回收调节蛋白EHD1的相互作用伙伴。我们通过GST-Pull、酵母双杂交实验、等温量热法和免疫共沉淀证明了这种相互作用，并发现EHD1的EH结构域与Rank-5的NPFED基序之间发生了结合。与EHD1类似，我们发现Rank-5与逆转录复合体的成分如Vps26共定位并相互作用，表明Rank-5在基于逆转录的运输中发挥作用。事实上，Rank-5的缺失会导致Vps26的错误定位，并影响甘露糖6-磷酸受体(M6PR)从内体运输到高尔基体的恢复和生物合成运输。此外，Rank-5对于正常的逆转录分子分布是必需的，因为野生型Rank-5-siRNA抗性结构的过度表达拯救了逆转录分子的错误定位。最后，我们发现Rank-5或EHD1的缺失会损害VSV-G的分泌。总体而言，我们的数据确定了Rank-5和EHD1之间的新相互作用，以及新的内分泌调节作用，包括基于逆转录的转运和分泌。
Rabankyrin-5 (Rank-5) has been implicated as an effector of the small GTPase Rab5 and plays an important role in macropinocytosis. We have now identified Rank-5 as an interaction partner for the recycling regulatory protein EHD1. We have demonstrated this interaction by GST-pulldown, yeast two-hybrid assay, isothermal calorimetry, and co-immunoprecipitation and found that the binding occurs between the EH-domain of EHD1 and the NPFED motif of Rank-5. Similar to EHD1, we found that Rank-5 co-localizes and interacts with components of the retromer complex such as Vps26, suggesting a role for Rank-5 in retromer-based transport. Indeed, depletion of Rank-5 causes mislocalization of Vps26 and affects both the retrieval of mannose 6-phosphate receptor (M6PR) transport to the Golgi from endosomes and biosynthetic transport. Moreover, Rank-5 is required for normal retromer distribution, as over-expression of a wild-type Rank-5-siRNA-resistant construct rescues retromer mislocalization. Finally, we show that depletion of either Rank-5 or EHD1 impairs secretion of VSV-G. Overall, our data identify a new interaction between Rank-5 and EHD1, and novel endocytic regulatory roles that include retromer-based transport and secretion.
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DOI：
10.1083/jcb.200408165
发表时间：
2004-11-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ang AL;Taguchi T;Francis S;Fölsch H;Murrells LJ;Pypaert M;Warren G;Mellman I
通讯作者：
Mellman I
影响因子：
4.5
作者：
Gokool, Suzanne;Tattersall, Daniel;Seaman, Matthew N. J.
通讯作者：
Seaman, Matthew N. J.
影响因子：
7.5
作者：
Bonifacino, Juan S.;Hurley, James H.
通讯作者：
Hurley, James H.
哺乳动物缩回术在与阳离子非依赖性甘露糖6-磷酸受体分类中的作用。
影响因子：
7.8
作者：
Arighi, Cecilia N;Hartnell, Lisa M;Aguilar, Ruben C;Haft, Carol R;Bonifacino, Juan S
通讯作者：
Bonifacino, Juan S
影响因子：
7.8
作者：
Caplan, S;Hartnell, L M;Aguilar, R C;Naslavsky, N;Bonifacino, J S
通讯作者：
Bonifacino, J S