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 nucleotide-dependent 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 of EHD2 on Rac1 activity was 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.
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Research Article|
October 13 2011
EHD2 mediates trafficking from the plasma membrane by modulating Rac1 activity
Sigi Benjamin;
Sigi Benjamin
1
*Department of Cell Research and Immunology, Tel Aviv University, Ramat Aviv 69978, Israel
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Hilla Weidberg;
Hilla Weidberg
2
*Department of Cell Research and Immunology, Tel Aviv University, Ramat Aviv 69978, Israel
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Debora Rapaport;
Debora Rapaport
*Department of Cell Research and Immunology, Tel Aviv University, Ramat Aviv 69978, Israel
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Olga Pekar;
Olga Pekar
*Department of Cell Research and Immunology, Tel Aviv University, Ramat Aviv 69978, Israel
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Marina Nudelman;
Marina Nudelman
*Department of Cell Research and Immunology, Tel Aviv University, Ramat Aviv 69978, Israel
†Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Ramat Aviv 69978, Israel
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Daniel Segal;
Daniel Segal
†Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Ramat Aviv 69978, Israel
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Koret Hirschberg;
Koret Hirschberg
‡Department of Pathology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv 69978, Israel
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Shulamit Katzav;
Shulamit Katzav
§The Hubert H. Humphrey Center for Experimental Medicine and Cancer Research, The Hebrew University Hadassah Medical School, Jerusalem, Israel
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Marcelo Ehrlich;
Marcelo Ehrlich
*Department of Cell Research and Immunology, Tel Aviv University, Ramat Aviv 69978, Israel
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Mia Horowitz
Mia Horowitz
3
*Department of Cell Research and Immunology, Tel Aviv University, Ramat Aviv 69978, Israel
3To whom correspondence should be addressed (email [email protected]).
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Publisher: Portland Press Ltd
Received:
June 07 2011
Accepted:
July 15 2011
Accepted Manuscript online:
July 15 2011
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2011 Biochemical Society
2011
Biochem J (2011) 439 (3): 433–445.
Article history
Received:
June 07 2011
Accepted:
July 15 2011
Accepted Manuscript online:
July 15 2011
Citation
Sigi Benjamin, Hilla Weidberg, Debora Rapaport, Olga Pekar, Marina Nudelman, Daniel Segal, Koret Hirschberg, Shulamit Katzav, Marcelo Ehrlich, Mia Horowitz; EHD2 mediates trafficking from the plasma membrane by modulating Rac1 activity. Biochem J 1 November 2011; 439 (3): 433–445. doi: https://doi.org/10.1042/BJ20111010
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