Accessory Protein Recruitment Motifs in Clathrin-Mediated Endocytosis

Brett, Tom J.; Traub, Linton M.; Fremont, Daved H.

doi:10.1016/s0969-2126(02)00784-0

Cited by 186 publications

(223 citation statements)

References 52 publications

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“…As seen in Figure 3B, EpsinR2C-13 bound to a-adaptin. Sequence analysis revealed that EpsinR2C-13 contains a DPF motif, which is the motif in animal epsin 1 that mediates binding to a-adaptin (Brett et al, 2002). To determine whether this motif was responsible for binding to a-adaptin, we generated an EpsinR2C-13 mutant in which the amino acids of the DPF motif were replaced with Ala (EpsinR2[DPF/AAA]).…”

Section: Epsinr2 Binds To A-adaptin and D-adaptinmentioning

confidence: 99%

EpsinR2 Interacts with Clathrin, Adaptor Protein-3, AtVTI12, and Phosphatidylinositol-3-Phosphate. Implications for EpsinR2 Function in Protein Trafficking in Plant Cells

et al. 2007

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Members of the epsin family of proteins (epsins) are characterized by the presence of an epsin N-terminal homology (ENTH) domain. Epsins have been implicated in various protein-trafficking pathways in animal and yeast (Saccharomyces cerevisiae) cells. Plant cells also contain multiple epsin-related proteins. In Arabidopsis (Arabidopsis thaliana), EPSIN1 is involved in vacuolar trafficking of soluble proteins. In this study, we investigated the role of Arabidopsis EpsinR2 in protein trafficking in plant cells. EpsinR2 contains a highly conserved ENTH domain but a fairly divergent C-terminal sequence. We found that the N-terminal ENTH domain specifically binds to phosphatidylinositol-3-P in vitro and has a critical role in the targeting of EpsinR2. Upon transient expression in protoplasts, hemagglutinin epitope-tagged EpsinR2 was translocated primarily to a novel cellular compartment, while a minor portion localized to the Golgi complex. Protein-binding experiments showed that EpsinR2 interacts with clathrin, AtVTI12, and the Arabidopsis homologs of adaptor protein-3 d-adaptin and adaptor protein-2 a-adaptin. Localization experiments revealed that hemagglutinin epitope-tagged EpsinR2 colocalizes primarily with d-adaptin and partially colocalizes with clathrin and AtVTI12. Based on these findings, we propose that EpsinR2 plays an important role in protein trafficking through interactions with d-adaptin, AtVTI12, clathrin, and phosphatidylinositol-3-P.

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Section: Epsinr2 Binds To A-adaptin and D-adaptinmentioning

confidence: 99%

EpsinR2 Interacts with Clathrin, Adaptor Protein-3, AtVTI12, and Phosphatidylinositol-3-Phosphate. Implications for EpsinR2 Function in Protein Trafficking in Plant Cells

et al. 2007

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“…We next searched for motifs that mediate binding to clathrin adaptor proteins and found that Ocrl contains an FxDxF AP-2 binding motif at residues 151-155 (Fig. 4a) but no F͞WXXF AP-1 binding motif (18)(19)(20). GST binding assays with GST-AP-2␣, GST-AP-1␥, and GST-GGA2 (170-613) show that purified Ocrl directly interacts with AP-2␣ but only slightly with either AP-1␥ or GGA2 (Fig.…”

Section: Ocrl Binds To the ␣ Appendage Subunit Of Ap-2 (Ap-2␣) And Comentioning

confidence: 99%

The inositol polyphosphate 5-phosphatase Ocrl associates with endosomes that are partially coated with clathrin

Ungewickell

Ward

Ungewickell

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

115

149

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The subcellular localization of Ocrl, the inositol polyphosphate 5-phosphatase that is mutated in Lowe syndrome, was investigated by fluorescence microscopy. Ocrl was localized to endosomes and Golgi membranes along with clathrin, giantin, the mannose 6-phosphate receptor, transferrin, and the early endosomal antigen 1 endosomal marker in fixed cells. The endosomal localization of Ocrl was confirmed by live-cell time-lapse microscopy in which we monitored the dynamics of Ocrl on endosomes. GST binding assays show that Ocrl interacts with the clathrin terminal domain and the clathrin adaptor protein AP-2. Our findings suggest a role for Ocrl in endosomal receptor trafficking and sorting.

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“…This tract is considerably longer than the DP(FW) (41)(42)(43), FXDXF (43), and WXX(FW)X(DE) (44 -46) sequence motifs that engage the alternative AP-2 appendage, the ␣ subunit appendage (Table I). To determine whether this ARH sequence can interact with AP-2 in isolation, we fused either GLDEAFSRLAQSRTNP (GST-ARHM1) or VWELDDG-LDEAFSRLAQSRTNP (GST-ARHM2) in-frame with GST.…”

Section: Glumentioning

confidence: 99%

Functional Dissection of an AP-2 β2 Appendage-binding Sequence within the Autosomal Recessive Hypercholesterolemia Protein

Mishra

Keyel

Edeling

et al. 2005

Journal of Biological Chemistry

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The autosomal recessive hypercholesterolemia (ARH) protein plays a critical role in regulating plasma low density lipoprotein (LDL) levels. Inherited defects in ARH lead to a hypercholesterolemia that closely phenocopies that caused by a defective LDL receptor. The elevated serum LDL-cholesterol levels typical of ARH patients and the pronounced accumulation of the LDL receptor at the cell surface of hepatocytes in ARH-null mice argue that ARH operates by promoting the internalization of the LDL receptor within clathrin-coated vesicles. ARH contains an amino-terminal phosphotyrosine-binding domain that associates physically with the LDL receptor internalization sequence and with phosphoinositides. The carboxyl-terminal half of ARH contains a clathrin-binding sequence and a separate AP-2 adaptor binding region providing a plausible mechanism for how ARH can act as an endocytic adaptor or CLASP (clathrin-associated sorting protein) to couple LDL receptors with the clathrin machinery. Because the interaction with AP-2 is highly selective for the independently folded appendage domain of the ␤2 subunit, we have characterized the ARH ␤2 appendagebinding sequence in detail. Unlike the known ␣ appendage-binding motifs, ARH requires an extensive sequence tract to bind the ␤ appendage with comparably high affinity. A minimal 16-residue sequence functions autonomously and depends upon ARH residues Asp 253 , Phe 259 , Leu 262 , and Arg 266 . We suggested that biased ␤ subunit engagement by ARH and the only other ␤2 appendage selective adaptor, ␤-arrestin, promotes efficient incorporation of this mechanistically distinct subset of CLASPs into clathrin-coated buds.Clathrin-coated vesicles are short lived transport intermediates fabricated solely for the preferential sorting and intracellular trafficking of select transmembrane proteins, bound ligands, and lipids (1-3). Clathrin coats assemble at several discrete intracellular sites where, typically, different cargo molecules are sorted. As the two major clathrin-associated heterotetrameric adaptor proteins (AP-1 and AP-2) are generally restricted to particular sorting stations within the cell, a long standing model has been that adaptors direct the process of cargo selection (4). Different subunits of the adaptor complex directly engage different classes of sorting sequences. The 2 subunit binds to YXXØ-type (where Ø is a bulky hydrophobic amino acid) sorting signals in a phosphorylation-dependent manner (3,5,6). In the basal state, the orientation of the 2 subunit within AP-2 makes the YXXØ interaction surface inaccessible. Phosphorylation on 2 residue Thr 156 is believed to alter the conformation of 2 to allow productive engagement of the YXXØ sequence (7-9). Another sorting sequence termed the (DE)XXXL(LI) motif is recognized by AP-1, AP-2, and AP-3 (6). Recent evidence suggests that the (DE)XXXL(LI) sequence is bound by a hemicomplex of the ␥ and 1 subunits in AP-1 or the ␦ and 3 subunits in AP-3 (10), although the precise location of the interaction site remains to be det...

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Accessory Protein Recruitment Motifs in Clathrin-Mediated Endocytosis

Cited by 186 publications

References 52 publications

EpsinR2 Interacts with Clathrin, Adaptor Protein-3, AtVTI12, and Phosphatidylinositol-3-Phosphate. Implications for EpsinR2 Function in Protein Trafficking in Plant Cells

EpsinR2 Interacts with Clathrin, Adaptor Protein-3, AtVTI12, and Phosphatidylinositol-3-Phosphate. Implications for EpsinR2 Function in Protein Trafficking in Plant Cells

The inositol polyphosphate 5-phosphatase Ocrl associates with endosomes that are partially coated with clathrin

Functional Dissection of an AP-2 β2 Appendage-binding Sequence within the Autosomal Recessive Hypercholesterolemia Protein

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