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Rabankyrin‐5 Interacts with EHD 1 and Vps 26 to Regulate Endocytic Trafficking and Retromer Function
Author(s) -
Zhang Jing,
Reiling Calliste,
Reinecke James B.,
Prislan Iztok,
Marky Luis A.,
Sorgen Paul L.,
Naslavsky Naava,
Caplan Steve
Publication year - 2012
Publication title -
traffic
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.677
H-Index - 130
eISSN - 1600-0854
pISSN - 1398-9219
DOI - 10.1111/j.1600-0854.2012.01334.x
Subject(s) - retromer , endosome , microbiology and biotechnology , endocytic cycle , biology , sorting nexin , transport protein , endocytosis , biochemistry , intracellular , receptor
Rabankyrin‐5 ( Rank ‐5) has been implicated as an effector of the small GTPase Rab 5 and plays an important role in macropinocytosis. We have now identified Rank ‐5 as an interaction partner for the recycling regulatory protein, Eps 15 homology domain 1 ( EHD 1). We have demonstrated this interaction by glutathione S‐transferase‐pulldown, yeast two‐hybrid assay, isothermal calorimetry and co‐immunoprecipitation, and found that the binding occurs between the EH domain of EHD 1 and the NPFED motif of Rank ‐5. Similar to EHD 1, we found that Rank ‐5 colocalizes and interacts with components of the retromer complex such as vacuolar protein sorting 26 ( Vps 26), suggesting a role for Rank ‐5 in retromer‐based transport. Indeed, depletion of Rank ‐5 causes mislocalization of Vps 26 and affects both the retrieval of mannose 6‐phosphate receptor transport to the Golgi from endosomes and biosynthetic transport. Moreover, Rank ‐5 is required for normal retromer distribution, as overexpression of a wild‐type Rank ‐5‐small interfering RNA ‐resistant construct rescues retromer mislocalization. Finally, we show that depletion of either Rank ‐5 or EHD 1 impairs secretion of vesicular stomatitis virus glycoprotein. Overall, our data identify a new interaction between Rank ‐5 and EHD 1, and novel endocytic regulatory roles that include retromer‐based transport and secretion.