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- Title
γ-Adaptin Interacts Directly with Rabaptin-5 through Its Ear Domain1.
- Authors
Shiba, Yoko; Takatsu, Hiroyuki; Shin, Hye-Won; Nakayama, Kazuhisa
- Abstract
In yeast two-hybrid screening using γ1-adaptin, a subunit of the AP-1 adaptor complex of clathrin-coated vesicles derived from the trans-Golgi network (TGN), as bait, we found that it could interact with Rabaptin-5, an effector of Rab5 and Rab4 that regulates membrane docking with endosomes. Further two-hybrid analysis revealed that the interaction occurs between the ear domain of γ1-adaptin and the COOH-terminal coiled-coil region of Rabaptin-5. Pull down assay with a fusion protein between glutathione S-transferase and the ear domain of γ1-adaptin and coimmunoprecipitation analysis revealed that the interaction occurs in vitro and in vivo. Immunocytochemical analysis showed that γ1-adaptin and Rabaptin-5 colocalize to a significant extent on perinuclear structures, probably on recycling endosomes, and are redistributed into the cytoplasm upon treatment with brefeldin A. These results suggest that the γ1-adaptin-Rabaptin-5 interaction may play a role in membrane trafficking between the TGN and endosomes.
- Subjects
RABAPTIN; ENDOSOMES; ORGANELLES; CYTOPLASM; CELLS
- Publication
Journal of Biochemistry, 2002, Vol 131, Issue 3, p327
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/oxfordjournals.jbchem.a003107