Rabankyrin-5 Interacts with EHD1 and Vps26 to Regulate Endocytic Trafficking and Retromer Function

Jing Zhang, Calliste Reiling, James B. Reinecke, Iztok Prislan, Luis A. Marky, Paul L. Sorgen, Naava Naslavsky, Steve Caplan

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

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, Eps15 homology domain 1 (EHD1). 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 EHD1 and the NPFED motif of Rank-5. Similar to EHD1, we found that Rank-5 colocalizes and interacts with components of the retromer complex such as vacuolar protein sorting 26 (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 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 EHD1 impairs secretion of vesicular stomatitis virus glycoprotein. Overall, our data identify a new interaction between Rank-5 and EHD1, and novel endocytic regulatory roles that include retromer-based transport and secretion.

Original languageEnglish (US)
Pages (from-to)745-757
Number of pages13
JournalTraffic
Volume13
Issue number5
DOIs
StatePublished - May 2012

Keywords

  • Biosynthetic transport
  • Golgi
  • Retromer
  • Secretion
  • VSV-G

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Fingerprint Dive into the research topics of 'Rabankyrin-5 Interacts with EHD1 and Vps26 to Regulate Endocytic Trafficking and Retromer Function'. Together they form a unique fingerprint.

  • Cite this