EHD2 mediates trafficking from the plasma membrane by modulating Rac1 activity

Sigi Benjamin, Hilla Weidberg, Debora Rapaport, Olga Pekar, Marina Nudelman, Daniel Segal, Koret Hirschberg, Shulamit Katzav, Marcelo Ehrlich, Mia Horowitz*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

EHDs [EH (Eps15 homology)-domain-containing proteins] participate in different stages of endocytosis. EHD2 is a plasma-membrane-associated EHD which regulates trafficking from the plasma membrane and recycling. EHD2 has a role in nucleotidedependent membrane remodelling and its ATP-binding domain is involved in dimerization, which creates a membrane-binding region. Nucleotide binding is important for association of EHD2 with the plasma membrane, since a nucleotide-free mutant (EHD2 T72A) failed to associate. To elucidate the possible function of EHD2 during endocytic trafficking, we attempted to unravel proteins that interact with EHD2, using the yeast two-hybrid system. A novel interaction was found between EHD2 and Nek3 [NIMA (never in mitosis in Aspergillus nidulans)-related kinase 3], a serine/threonine kinase. EHD2 was also found in association with Vav1, a Nek3-regulated GEF (guanine-nucleotide- exchange factor) for Rho GTPases. Since Vav1 regulates Rac1 activity and promotes actin polymerization, the impact of overexpression ofEHD2on Rac1 activitywas tested. The results indicated that wt (wild-type) EHD2, but not its P-loop mutants, reduced Rac1 activity. The inhibitory effect of EHD2 overexpression was partially rescued by co-expression of Rac1 as measured using a cholera toxin trafficking assay. The results of the present study strongly indicate that EHD2 regulates trafficking from the plasma membrane by controlling Rac1 activity.

Original languageEnglish
Pages (from-to)433-442
Number of pages10
JournalBiochemical Journal
Volume439
Issue number3
DOIs
StatePublished - 1 Nov 2011

Keywords

  • Endocytosis
  • Eps15 homology (EH)-domain-containing protein 2 (EHD2)
  • Rac1 activity

Fingerprint

Dive into the research topics of 'EHD2 mediates trafficking from the plasma membrane by modulating Rac1 activity'. Together they form a unique fingerprint.

Cite this