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Chemical-genetic disruption of clathrin function spares adaptor complex 3-dependent endosome vesicle biogenesis.

Molecular biology of the cell | 2013

A role for clathrin in AP-3-dependent vesicle biogenesis has been inferred from biochemical interactions and colocalization between this adaptor and clathrin. The functionality of these molecular associations, however, is controversial. We comprehensively explore the role of clathrin in AP-3-dependent vesicle budding, using rapid chemical-genetic perturbation of clathrin function with a clathrin light chain-FKBP chimera oligomerizable by the drug AP20187. We find that AP-3 interacts and colocalizes with endogenous and recombinant FKBP chimeric clathrin polypeptides in PC12-cell endosomes. AP-3 displays, however, a divergent behavior from AP-1, AP-2, and clathrin chains. AP-3 cofractionates with clathrin-coated vesicle fractions isolated from PC12 cells even after clathrin function is acutely inhibited by AP20187. We predicted that AP20187 would inhibit AP-3 vesicle formation from endosomes after a brefeldin A block. AP-3 vesicle formation continued, however, after brefeldin A wash-out despite impairment of clathrin function by AP20187. These findings indicate that AP-3-clathrin association is dispensable for endosomal AP-3 vesicle budding and suggest that endosomal AP-3-clathrin interactions differ from those by which AP-1 and AP-2 adaptors productively engage clathrin in vesicle biogenesis.

Pubmed ID: 23761069 RIS Download

Associated grants

  • Agency: NINDS NIH HHS, United States
    Id: R56 NS042599
  • Agency: NIEHS NIH HHS, United States
    Id: T32 ES012870
  • Agency: NINDS NIH HHS, United States
    Id: F31NS0765
  • Agency: NIGMS NIH HHS, United States
    Id: GM077569
  • Agency: NIEHS NIH HHS, United States
    Id: P01ES016731
  • Agency: NIGMS NIH HHS, United States
    Id: R01 GM077569
  • Agency: NIEHS NIH HHS, United States
    Id: T32ES012870
  • Agency: NINDS NIH HHS, United States
    Id: P30NS055077
  • Agency: NIGMS NIH HHS, United States
    Id: T32 GM008169
  • Agency: NINDS NIH HHS, United States
    Id: NS42599
  • Agency: NIEHS NIH HHS, United States
    Id: P01 ES016731
  • Agency: NINDS NIH HHS, United States
    Id: R01 NS042599
  • Agency: Medical Research Council, United Kingdom
    Id: G120/952
  • Agency: NINDS NIH HHS, United States
    Id: P30 NS055077

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