The Interaction of Epsin and Eps15 with the Clathrin Adaptor AP-2 Is Inhibited by Mitotic Phosphorylation and Enhanced by Stimulation-dependent Dephosphorylation in Nerve Terminals

Chen, Hong; Slepnev, Vladimir I.; Fiore, Pier Paolo Di; Camilli, Pietro De

doi:10.1074/jbc.274.6.3257

Cited by 130 publications

(108 citation statements)

References 30 publications

Supporting

Mentioning

101

Contrasting

Order By: Relevance

“…As we have shown previously, some of the interactions of epsin are regulated by its phosphorylation and reversible ubiquitination (15,46). Here, we report evidence for an additional control mechanism based on its binding to clathrin.…”

Section: Discussionsupporting

confidence: 78%

The association of epsin with ubiquitinated cargo along the endocytic pathway is negatively regulated by its interaction with clathrin

Chen

Camilli

2005

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

Self Cite

137

142

View full text Add to dashboard Cite

Monoubiquitination of plasma membrane proteins is a mechanism to control their endocytic trafficking by promoting their interaction with cytosolic adaptor proteins that contain ubiquitin (Ub)-binding domains. Epsin, which contains Ub interaction motifs (UIMs), as well as binding sites for the clathrin coat and clathrin accessory factors, is thought to function as one of such adaptors. The importance of clathrin in the internalization of ubiquitinated cargo, however, has been questioned. Here, we show that a GFP-Ub chimera directly targeted to the plasma membrane via a lipidbased interaction is efficiently taken up by endocytosis and delivered to the same endosomes that accumulate internalized EGF. Internalization of the chimera requires integrity of the UIM binding interface of Ub, but does not require clathrin. Surprisingly, WT epsin showed little colocalization with this chimera, whereas UIM-containing epsin constructs that lack the clathrin and AP2 binding region, strikingly colocalized with this chimera on endocytic vacuoles. In addition, extensive colocalization of WT epsin with the chimera on endocytic structures could be observed in cells where clathrin levels were drastically reduced by RNA interference. Our results reveal an important regulatory mechanism in epsin function. The mutually exclusive colocalization of epsin with membrane-bound Ub or clathrin may play a role in controlling the endocytic route taken by ubiquitinated cargo.ubiquitin ͉ endocytosis ͉ Rab5 ͉ endosome ͉ FYVE domain P rotein ubiquitination plays pleotropic roles in cell physiology.

show abstract

Section: Discussionsupporting

confidence: 78%

The association of epsin with ubiquitinated cargo along the endocytic pathway is negatively regulated by its interaction with clathrin

Chen

Camilli

2005

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

Self Cite

137

142

View full text Add to dashboard Cite

show abstract

“…EPS15 was first identified as a substrate of EGFR kinase (56), but was subsequently found to participate in clathrin-mediated endocytosis through interaction with the clathrin adaptor protein AP2, and could mediate clathrin-coated pit formation (57). The DN mutant of EPS15 decreased HLHCGR internalization and in addition led to increased agonist-stimulated cAMP production.…”

Section: )mentioning

confidence: 99%

ARF6 Activated by the LHCG Receptor through the Cytohesin Family of Guanine Nucleotide Exchange Factors Mediates the Receptor Internalization and Signaling

Kanamarlapudi

Thompson

Kelly

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background:The mechanisms by which ARF6 regulate LHCGR internalization and signaling are unknown. Results: Heterotrimeric G-protein, PI 3-kinase, cytohesin ARF GEFs and ARF GAPs function upstream whereas dynamin and clathrin act downstream of ARF6 in the regulation of HLHCGR internalization and signaling. Conclusion: Dissected the molecular mechanisms underlying ARF6 involvement in LHCGR internalization and signaling. Significance: This study provides insight into the mechanisms responsible for HLHCGR regulation by ARF6.

show abstract

“…Dephosphorylation of components of endocytic machinery also appears to be necessary in vivo. When calcium enters the nerve terminal, calcineurin triggers the dephosphorylation of dynamin, amphiphysin, synaptojanin, epsin, and eps15, allowing them to assemble with AP-2 into a macromolecular complex, which presumably is required for the internalization of plasma membrane (Bauerfeind et al, 1997;Slepnev et al, 1998;Chen et al, 1999). The dephosphorylation of ␤-arrestin also is required for its association with clathrin (Lin et al, 1997).…”

Section: Ap-3 Adaptor Phosphorylation By Casein Kinasementioning

confidence: 99%

The AP-3 Complex Required for Endosomal Synaptic Vesicle Biogenesis is Associated with a Casein Kinase Ια-Like Isoform

Faúndez

Kelly

2000

MBoC

View full text Add to dashboard Cite

The formation of small vesicles is mediated by cytoplasmic coats the assembly of which is regulated by the activity of GTPases, kinases, and phosphatases. A heterotetrameric AP-3 adaptor complex has been implicated in the formation of synaptic vesicles from PC12 endosomes . When the small GTPase ARF1 is prevented from hydrolyzing GTP, we can reconstitute AP-3 recruitment to synaptic vesicle membranes in an assembly reaction that requires temperatures above 15°C and the presence of ATP suggesting that an enzymatic step is involved in the coat assembly. We have now found an enzymatic reaction, the phosphorylation of the AP-3 adaptor complex, that is linked with synaptic vesicle coating. Phosphorylation occurs in the ␤3 subunit of the complex by a kinase similar to casein kinase 1␣. The kinase copurifies with neuronal-specific AP-3. In vitro, purified casein kinase I selectively phosphorylates the ␤3A and ␤3B subunit at its hinge domain. Inhibiting the kinase hinders the recruitment of AP-3 to synaptic vesicles. The same inhibitors that prevent coat assembly in vitro also inhibit the formation of synaptic vesicles in PC12 cells. The data suggest, therefore, that the mechanism of AP-3-mediated vesiculation from neuroendocrine endosomes requires the phosphorylation of the adaptor complex at a step during or after AP-3 recruitment to membranes. INTRODUCTIONMembrane proteins are carried from donor membranes to acceptor membranes by two steps, the vesiculation of a carrier vesicle from the donor and the fusion of the carrier vesicle with the acceptor. Vesiculation of the donor membrane can itself be divided into five steps: recognition of cargo proteins; recruitment of coating molecules; the imposition of curvature on the forming vesicle membrane; fission of the carrier vesicle from the donor membrane; and, finally, the loss of the vesicle coat (Springer et al., 1999). To discover the molecular events that take place at each step, several laboratories have reconstituted either the entire process of vesiculation or individual steps in vitro.The first two steps of vesiculation, cargo recognition and coating, can be regulated by phosphorylation. Membrane proteins are recognized as cargo because they contain sorting domains. Adaptors interact with a tyrosine or a dileucine motif (Schmid, 1997;Kirchhausen et al., 1997;Bonifacino and Dell'Angelica, 1999). The recognition of cargo molecules can be regulated by the phosphorylation of sites close to the sorting domains; for example, in the trafficking of furin (Wan et al., 1998;Teuchert et al., 1999;Molloy et al., 1999), CD4 (Pitcher et al., 1999), CTLA-4 (Shiratori et al., 1997), MHC-II-Ii complex (Anderson and Roche, 1998), and pIgAR (Casanova et al., 1990;Okamoto et al., 1994;Luton et al., 1998). The cargo molecules are recruited into a newly forming carrier vesicle by cytoplasmic coat molecules (Matsuoka et al., 1998;Bremser et al., 1999). The recruitment of two of the known coats, COPI and COPII, is regulated by small ARF-like GTPases and appears to be a relatively simple p...

show abstract

The Interaction of Epsin and Eps15 with the Clathrin Adaptor AP-2 Is Inhibited by Mitotic Phosphorylation and Enhanced by Stimulation-dependent Dephosphorylation in Nerve Terminals

Cited by 130 publications

References 30 publications

The association of epsin with ubiquitinated cargo along the endocytic pathway is negatively regulated by its interaction with clathrin

The association of epsin with ubiquitinated cargo along the endocytic pathway is negatively regulated by its interaction with clathrin

ARF6 Activated by the LHCG Receptor through the Cytohesin Family of Guanine Nucleotide Exchange Factors Mediates the Receptor Internalization and Signaling

The AP-3 Complex Required for Endosomal Synaptic Vesicle Biogenesis is Associated with a Casein Kinase Ια-Like Isoform

Contact Info

Product

Resources

About