.GAMMA.-Adaptin Interacts Directly with Rabaptin-5 through Its Ear Domain.

J-STAGE
  • Shiba Yoko
    Institute of Biological Sciences and Gene Research Center, University of Tsukuba, Tsukuba Science City
  • Takatsu Hiroyuki
    Institute of Biological Sciences and Gene Research Center, University of Tsukuba, Tsukuba Science City
  • Shin Hye-Won
    Institute of Biological Sciences and Gene Research Center, University of Tsukuba, Tsukuba Science City
  • Nakayama Kazuhisa
    Institute of Biological Sciences and Gene Research Center, University of Tsukuba, Tsukuba Science City

Description

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 coiledcoil 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.

Journal

  • J Biochem (Tokyo)

    J Biochem (Tokyo) 131 (3), 327-336, 2002

    The Japanese Biochemical Society

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