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Antibody to AP1B Adaptor Blocks Biosynthetic and Recycling Routes of Basolateral Proteins at Recycling Endosomes

Cancino, Jorge, Torrealba, Carolina, Soza, Andrea, Yuseff, María Isabel, Gravotta, Diego, Henklein, Peter, Rodriguez-Boulan, Enrique, González, Alfonso·2007·Molecular Biology of the Cell
DOI 10.1091/mbc.e07-06-0563
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Abstract

The epithelial-specific adaptor AP1B sorts basolateral plasma membrane (PM) proteins in both biosynthetic and recycling routes, but the site where it carries out this function remains incompletely defined. Here, we have investigated this topic in Fischer rat thyroid (FRT) epithelial cells using an antibody against the medium subunit μ1B. This antibody was suitable for immunofluorescence and blocked the function of AP1B in these cells. The antibody blocked the basolateral recycling of two basolateral PM markers, Transferrin receptor (TfR) and LDL receptor (LDLR), in a perinuclear compartment with marker and functional characteristics of recycling endosomes (RE). Live imaging experiments demonstrated that in the presence of the antibody two newly synthesized GFP-tagged basolateral proteins (vesicular stomatitis virus G [VSVG] protein and TfR) exited the trans-Golgi network (TGN) normally but became blocked at the RE within 3–5 min. By contrast, the antibody did not block trafficking of green fluorescent protein (GFP)-LDLR from the TGN to the PM but stopped its recycling after internalization into RE in ∼45 min. Our experiments conclusively demonstrate that 1) AP1B functions exclusively at RE; 2) TGN-to-RE transport is very fast and selective and is mediated by adaptors different from AP1B; and 3) the TGN and AP1B-containing RE cooperate in biosynthetic basolateral sorting.

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Metadata

Authors
Cancino, J., Torrealba, C., Soza, A., Yuseff, M. I., Gravotta, D., Henklein, P., Rodriguez-Boulan, E., & González, A.
Affiliations
  • *Departamento de Inmunología Clínica y Reumatología, Facultad de Medicina, and Centro de Regulación Celular y Patología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, 6510260 Santiago, Chile;
  • Dyson Vision Research Institute, Weill Medical College of Cornell University, New York, NY 10021
  • Institute of Biochemistry Faculty of Medicine, Humboldt University, 10117 Berlin, Germany; and
Venue
Molecular Biology of the Cell
Publisher
American Society for Cell Biology (ASCB)
Published
2007-12
Volume
18
Issue
12
Pages
4872-4884
ISSN
  • 1059-1524
  • 1939-4586
DOI
10.1091/mbc.e07-06-0563
Cited by
91
References
0
Language
en

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