100-kDa polypeptides in peripheral clathrin-coated vesicles are required for receptor-mediated endocytosis.

Chin, Daniel J.; Straubinger, Robert M.; Acton, Susan; Näthke, Inke; Brodsky, Frances M.

doi:10.1073/pnas.86.23.9289

Cited by 104 publications

(62 citation statements)

References 26 publications

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“…This interaction might potentiate receptor migration, association with coated pits, and targeting to endosomes. Such a function is supported by the observation that microinjection of an anti-a-adaptin monoclonal antibody into HeLa cells inhibits the down regulation of the transferrin receptor, resulting in the maintenance of the newly endocytosed receptors in a peripheral cytoplasmic distribution rather than a perinuclear distribution (5). Analysis of the role of a-adaptin in receptor internalization has been primarily limited to receptors, such as the low-density lipoprotein and transferrin receptors, which do not have intrinsic kinase activity.…”

Section: Discussionmentioning

confidence: 80%

The conserved C-terminal domain of the bovine papillomavirus E5 oncoprotein can associate with an alpha-adaptin-like molecule: a possible link between growth factor receptors and viral transformation.

Cohen¹,

Lowy²,

Schiller³

1993

Mol. Cell. Biol.

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The bovine papillomavirus E5 gene encodes an oncoprotein that can independently transform rodent fibroblasts. This small 44-amino-acid protein is thought to function through the activation of growth factor receptors. E5 activation of the epidermal growth factor receptor results in an increase in the number of activated receptors at the cell surface. This finding suggests that E5 may act by inhibiting the normal down regulation of activated epidermal growth factor receptor via coated pit-mediated endocytosis. We have constructed a fusion protein consisting of glutathione S-transferase and the conserved C-terminal domain of E5 (GST-E5) in order to identify E5-associated cellular proteins that may be involved in its transforming activity.We have identified a 125-kDa cellular protein with a strong associated serine kinase activity that specifically associated with GST-E5 in the reduced form but not with GST-E5 fusions that contained changes in several conserved amino acids. Microsequence and biochemical analyses suggest that p125 is a novel member of the a-adaptin family. Since ce-adaptins have previously been shown to be involved in coated pit-mediated cell surface receptor endocytosis and down regulation, these results suggest that p125 may be an a-adaptin-like molecule involved in growth factor receptor down regulation and that E5 may act by inhibiting its activity.The bovine papillomavirus E5 gene is the smallest known oncogene and is the major transforming gene within the bovine papillomavirus genome (23). It encodes a 44-aminoacid, membrane-associated protein that forms homodimers via two cysteines located near the C terminus (2). The E5 monomer is composed of an N-terminal 30-amino-acid hydrophobic transmembrane domain and a C-terminal 14-amino-acid hydrophilic domain (24). The hydrophilic domain contains a number of amino acids that are conserved among the papillomaviruses that induce fibropapillomas. The conserved residues, which include the cysteines required for homodimer formation, are important for E5-mediated transformation of rodent fibroblasts (10).E5 can transform cells via potentiation of the epidermal growth factor receptors (EGF-R) (14) or platelet-derived growth factor receptors (PDGF-R) (18). However, E5 may stimulate the activity of these two types of receptors by different mechanisms (6). E5 activates immature forms of the PDGF-R in the trans-Golgi through a ligand-independent mechanism, while there is no evidence for activation of immature forms of the EGF-R. PDGF-R stimulation appears to require dimerization of E5; this form of E5 is found in a complex with the PDGF-R and a 16-kDa protein that has been identified as a subunit of a vacuolar H+ ATPase (16K) (8). These interactions appear to be mediated by the transmembrane domains of E5 and the PDGF-R (6, 7). By contrast, potentiation of EGF-R activity by E5 seems to involve an inhibition of the normal down regulation of activated receptors in that increased numbers of activated EGF-R are detected at the cell surface, both in the presence and...

show abstract

Section: Discussionmentioning

confidence: 80%

The conserved C-terminal domain of the bovine papillomavirus E5 oncoprotein can associate with an alpha-adaptin-like molecule: a possible link between growth factor receptors and viral transformation.

Cohen¹,

Lowy²,

Schiller³

1993

Mol. Cell. Biol.

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“…Since the adaptors appear to recycle back through the soluble pool to the plasma membrane there must therefore be a second round ofuncoating, the site and mechanism of which has not yet been identified. However, adaptors have been detected on early endosomes (Chin et al, 1989) suggesting that either endocytic vesicles can fuse with early endosomes while still coated with adaptors or that only partial uncoating is necessary for fusion to occur.…”

Section: Uncout Ingmentioning

confidence: 99%

The mechanism of receptor‐mediated endocytosis

Smythe

Warren

1991

European Journal of Biochemistry

153

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“…Goat anti-clathrin heavy chain antibodies (Sigma-Aldrich, St. Louis, MO), anti-clathrin heavy chain monoclonal antibodies (mAbs) clone 23 (BD Biosciences, San Jose, CA), X22 (Chin et al, 1989), and rabbit polyclonal clathrin light-chain antibody 4878 (generously provided by S. Schmid, The Scripps Research Institute, La Jolla, CA) were used at dilutions of 1:100 -1:250. Anti-␣-adaptin (AP-2 subunit) mAbs (clone AP6; Affinity Bioreagents, Golden, CO) were used at a dilution of 1:200.…”

Section: Antibodies and Staining Reagentsmentioning

confidence: 99%

Internalization of Large Double-Membrane Intercellular Vesicles by a Clathrin-dependent Endocytic Process

Piehl

Lehmann

Gumpert

et al. 2007

MBoC

156

233

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Beyond its well-documented role in vesicle endocytosis, clathrin has also been implicated in the internalization of large particles such as viruses, pathogenic bacteria, and even latex beads. We have discovered an additional clathrin-dependent endocytic process that results in the internalization of large, double-membrane vesicles at lateral membranes of cells that are coupled by gap junctions (GJs). GJ channels bridge apposing cell membranes to mediate the direct transfer of electrical currents and signaling molecules from cell to cell. Here, we report that entire GJ plaques, clusters of GJ channels, can be internalized to form large, double-membrane vesicles previously termed annular gap junctions (AGJs). These internalized AGJ vesicles subdivide into smaller vesicles that are degraded by endo/lysosomal pathways. Mechanistic analyses revealed that clathrin-dependent endocytosis machinery-components, including clathrin itself, the alternative clathrin-adaptor Dab2, dynamin, myosin-VI, and actin are involved in the internalization, inward movement, and degradation of these large, intercellular double-membrane vesicles. These findings contribute to the understanding of clathrin's numerous emerging functions. INTRODUCTIONThe role of clathrin in endocytosis is well documented. This protein forms a typical curved lattice around endocytic vesicles that are internalized at the plasma membrane (PM). In addition, the involvement of clathrin in several uncharacteristic endocytic processes has been reported, including the internalization of viruses, pathogenic bacteria, and large latex beads (Aggeler and Werb, 1982;Ehrlich et al., 2004;Rust et al., 2004;Veiga and Cossart, 2005). Here, we describe another function of the clathrin-dependent endocytic machinery that results in the internalization of large, doublemembrane vesicles at lateral PMs of cells that are coupled by gap junctions (GJs).GJs are ubiquitously distributed channels that connect the cytoplasms of two apposing cells each participating in this connection via a half channel termed a connexon to provide direct cell-to-cell communication. Connexons are hexamers of four-pass membrane proteins called connexins (Cxs;Bruzzone et al., 1996;Kumar and Gilula, 1996). Once transported to the PM, GJ channels cluster into two-dimensional arrays termed plaques that can be composed of a few to many thousands of individual channels and vary from a few square nanometers to many square micrometers (Bruzzone et al., 1996; Falk, 2000a;Severs et al., 2001). GJ channels can open and close (gate) and physiological parameters, including intracellular pH, Ca 2ϩ concentration, and Cx phosphorylation, are known to modulate GJ channel gating and the extent of GJ-mediated intercellular coupling (Delmar et al., 2004;Lampe and Lau, 2004;Moreno, 2005). However, the extent of intercellular coupling could also be regulated through altering the number of GJ channels in the PM.Cxs have a surprisingly short half-life of only 1-5 h, leading to a rapid GJ and Cx protein turnover (Fallon and Goodeno...

show abstract

100-kDa polypeptides in peripheral clathrin-coated vesicles are required for receptor-mediated endocytosis.

Cited by 104 publications

References 26 publications

The conserved C-terminal domain of the bovine papillomavirus E5 oncoprotein can associate with an alpha-adaptin-like molecule: a possible link between growth factor receptors and viral transformation.

The conserved C-terminal domain of the bovine papillomavirus E5 oncoprotein can associate with an alpha-adaptin-like molecule: a possible link between growth factor receptors and viral transformation.

The mechanism of receptor‐mediated endocytosis

Internalization of Large Double-Membrane Intercellular Vesicles by a Clathrin-dependent Endocytic Process

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