期刊
TRAFFIC
卷 4, 期 11, 页码 802-811出版社
WILEY
DOI: 10.1034/j.1600-0854.2003.00136.x
关键词
AP-1; AP-3; BFA; endosomes; MPR; TGN
类别
The mannose 6-phosphate receptor MPR46 mediates sorting of lysosomal enzymes and recycles between the trans-Golgi network and endosomes. We characterized the retrograde transport of MPR46 from endosomes to the TGN by an in vitro transport assay using mouse fibroblast cell lines. Sulfation of a modified MPR46 upon entering the TGN is measured. The in vitro retrograde transport is time-, temperature-, ATP- and cytosol-dependent. Transport requires the SNARE proteins Vti1a and Syntaxin 16 and the Rab family member Rab6. The transport is sensitive to GTPgammaS, brefeldin A and independent of TIP47. These data indicate that MPR46 follows an early endosome-to-TGN route. Transport is inhibited by MPR46 tail peptide comprising the acidic cluster-di-leucine sorting motif to which adaptor proteins AP-1 and AP-3 bind. Transport depends on cytosolic AP-3, but not on cytosolic AP-1. Residual membrane-associated AP-1 may have masked a requirement for cytosolic AP-1. The competence of membranes from AP-1-deficient cells for endosome-to-TGN transport in vitro was severely compromised.
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