ARF1.GTP, tyrosine-based signals, and phosphatidylinositol 4,5-bisphosphate constitute a minimal machinery to recruit the AP-1 clathrin adaptor to membranes
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
 
ID 153656
Author(s) Crottet, P.; Meyer, D. M.; Rohrer, J.; Spiess, M.
Author(s) at UniBasel Spiess, Martin
Year 2002
Title ARF1.GTP, tyrosine-based signals, and phosphatidylinositol 4,5-bisphosphate constitute a minimal machinery to recruit the AP-1 clathrin adaptor to membranes
Journal Molecular Biology of the Cell
Volume 13
Number 10
Pages / Article-Number 3672-3682
Keywords ADP-Ribosylation Factor 1/isolation & purification/*metabolism; Adaptor Protein Complex 1/isolation & Amino Acid Sequence; Animals; Cattle; Cell Membrane/metabolism; Clathrin-Coated Vesicles/metabolism; Guanosine Triphosphate/chemistry/*metabolism; Lipid Metabolism; Lipids/chemistry; Liposomes/chemistry/metabolism; Molecular Sequence Data; Molecular Structure; Peptides/genetics/metabolism; Phosphatidylinositol 4; 5-Diphosphate/*metabolism; Protein Transport/physiology; Sequence Alignment; Signal Transduction/*physiology; Tyrosine/*metabolism; trans-Golgi Network/metabolism
Abstract At the trans-Golgi network, clathrin coats containing AP-1 adaptor complexes are formed in an ARF1-dependent manner, generating vesicles transporting cargo proteins to endosomes. The mechanism of site-specific targeting of AP-1 and the role of cargo are poorly understood. We have developed an in vitro assay to study the recruitment of purified AP-1 adaptors to chemically defined liposomes presenting peptides corresponding to tyrosine-based sorting motifs. AP-1 recruitment was found to be dependent on myristoylated ARF1, GTP or nonhydrolyzable GTP-analogs, tyrosine signals, and small amounts of phosphoinositides, most prominently phosphatidylinositol 4,5-bisphosphate, in the absence of any additional cytosolic or membrane bound proteins. AP-1 from cytosol could be recruited to a tyrosine signal independently of the lipid composition, but the rate of recruitment was increased by phosphatidylinositol 4,5-bisphosphate. The results thus indicate that cargo proteins are involved in coat recruitment and that the local lipid composition contributes to specifying the site of vesicle formation.
Publisher American Society for Cell Biology
ISSN/ISBN 1059-1524 ; 1939-4586
edoc-URL http://edoc.unibas.ch/dok/A5258051
Full Text on edoc Available
Digital Object Identifier DOI 10.1091/mbc.E02-05-0309
Document type (ISI) Article
 
   

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