title: Disabled-2 exhibits the properties of a cargo-selective endocytic clathrin adaptor.
abstract: Clathrin-coated pits at the cell surface select material for transportation into the cell interior. A major mode of cargo selection at the bud site is via the micro 2 subunit of the AP-2 adaptor complex, which recognizes tyrosine-based internalization signals. Other internalization motifs and signals, including phosphorylation and ubiquitylation, also tag certain proteins for incorporation into a coated vesicle, but the mechanism of selection is unclear. Disabled-2 (Dab2) recognizes the FXNPXY internalization motif in LDL-receptor family members via an N-terminal phosphotyrosine-binding (PTB) domain. Here, we show that in addition to binding AP-2, Dab2 also binds directly to phosphoinositides and to clathrin, assembling triskelia into regular polyhedral coats. The FXNPXY motif and phosphoinositides contact different regions of the PTB domain, but can stably anchor Dab2 to the membrane surface, while the distal AP-2 and clathrin-binding determinants regulate clathrin lattice assembly. We propose that Dab2 is a typical member of a growing family of cargo-specific adaptor proteins, including beta-arrestin, AP180, epsin, HIP1 and numb, which regulate clathrin-coat assembly at the plasma membrane by synchronizing cargo selection and lattice polymerization events.
authors.affiliation: Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, 3500 Terrace Street, S325BST, Pittsburgh, PA 15261, USA.
authors.forename: Sanjay K
authors.initials: SK
authors.lastname: Mishra
authors.forename: Peter A
authors.initials: PA
authors.lastname: Keyel
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Hawryluk
authors.forename: Nicole R
authors.initials: NR
authors.lastname: Agostinelli
authors.forename: Simon C
authors.initials: SC
authors.lastname: Watkins
authors.forename: Linton M
authors.initials: LM
authors.lastname: Traub
mesh_terms: D033962:Adaptor Protein Complex 2 / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D001692:Biological Transport / Q000502:physiology; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism* / Q000648:ultrastructure; D004705:Endocytosis / Q000502:physiology*; D005456:Fluorescent Dyes / Q000378:metabolism; D017930:Genes, Reporter; D016147:Genes, Tumor Suppressor; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011973:Receptors, LDL / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins; D033921:Vesicular Transport Proteins
chemical_list: D033962:Adaptor Protein Complex 2; D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins; D002966:Clathrin; C102885:DAB2 protein, human; D005456:Fluorescent Dyes; D009479:Neuropeptides; D010455:Peptides; D010716:Phosphatidylinositols; D011506:Proteins; D011973:Receptors, LDL; D011993:Recombinant Fusion Proteins; D025521:Tumor Suppressor Proteins; D033921:Vesicular Transport Proteins; C114154:epsin