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Transport of cholera toxin B-subunit from recycling endosomes to the Golgi requires clathrin and AP-1

Matsudaira, Tatsuyuki, Niki, Takahiro, Taguchi, Tomohiko, Arai, Hiroyuki·2015·Journal of Cell Science

Abstract

The retrograde pathway is defined by the transport of proteins and lipids from the plasma membrane (PM) through endosomes to the Golgi, and is essential for a variety of cellular activities. Recycling endosomes (REs) are important sorting stations for some retrograde cargo. SMAP2, an Arf GTPase-activating protein (GAP) for Arf1, with a putative clathrin-binding domain was previously shown to participate in the retrograde transport of CTxB from REs. Here we found that clathrin, a vesicle coat protein, and clathrin adaptor protein complex 1 (AP-1) were present at REs and were needed for the retrograde transport of cholera toxin B subunit (CTxB) from REs to the Golgi, but not from the PM to REs. SMAP2 immunoprecipitated clathrin/AP-1 through a putative clathrin-binding domain and a CALM-binding domain, and SMAP2 mutants that did not interact with clathrin/AP-1 could not localize to REs. Moreover, knockdown of Arf1 suppressed the retrograde transport of CTxB from REs to the Golgi. These findings suggest a clathrin-coated-vesicles-mediated retrograde transport from REs and the role of the coat proteins in the recruitment of Arf GAP to transport vesicles.

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  • AP-1 localizes to Rab11-positive recycling endosomes.

    “Since AP-1 has been shown to localize to Rab11-positive recycling endosomes [28], and ligand-stimulated EGFR is recycled back to the PM through Rab11-positive endosomes [29, 30], we examined if three molecules, EGFR, AP-1, and Rab11, could show colocalization in the cytoplasm.”
  • AP-1 localizes to Rab11-positive recycling endosomes, promoting retrograde transport of cholera toxin B-subunit from endosomes to the Golgi complex.

    “AP-1 has been shown to localize to Rab11-positive recycling endosomes, thereby promoting retrograde transport of the cholera toxin B-subunit from endosomes to the Golgi complex [28].”

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Metadata

Authors
Matsudaira, T., Niki, T., Taguchi, T., & Arai, H.
Affiliations
Department of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, Tokyo, 113-0033, Japan
Venue
Journal of Cell Science
Published
2015-01-01
ISSN
  • 1477-9137
  • 0021-9533
Cited by
22
References
3
Language
en

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