Clathrin functions in the absence of heterotetrameric adaptors and AP180-related proteins in yeast

Huang, Kristen M.; D'Hondt, Kathleen; Riezman, Howard; Lemmon, Sandra K.

doi:10.1093/emboj/18.14.3897

Cited by 133 publications

(104 citation statements)

References 100 publications

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“…Consistently, AP-2 was found not to be required for direct interaction with the EGFR for clathrin-dependent internalization of the EGFR (Huang et al, 1999;Nesterov et al, 1999). Furthermore, it was recently reported that the EGFR was efficiently endocytosed even in the absence of AP-2 upon RNA interference knock-down (Conner and Schmid, 2003;Motley et al, 2003).…”

Section: Discussionmentioning

confidence: 77%

Cbl-dependent Ubiquitination Is Required for Progression of EGF Receptors into Clathrin-coated Pits

Stang

Blystad

Kazazic

et al. 2004

MBoC

146

168

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Ligand binding causes the EGF receptor (EGFR) to become ubiquitinated by Cbl upon association with the adaptor protein Grb2. We have investigated the role of ubiquitin and Grb2 in ligand-induced endocytosis of the EGFR. Incubation of cells with EGF on ice caused translocation of Grb2 and Cbl from the cytosol to the rim of coated pits. Grb2 with point mutations in both SH3 domains inhibited recruitment of the EGFR to clathrin-coated pits, in a Ras-independent manner. On overexpression of the Cbl-binding protein Sprouty, ubiquitination of the EGFR was inhibited, the EGFR was recruited only to the rim of coated pits, and endocytosis of the EGFR was inhibited. Conjugation-defective ubiquitin similarly inhibited recruitment of EGF-EGFR to clathrin-coated pits. Even though this does not prove that cargo must be ubiquitinated, this indicates the importance of interaction of ubiquitinated protein(s) with proteins harboring ubiquitininteracting domains. We propose that Grb2 mediates transient anchoring of the EGFR to an Eps15-containing molecular complex at the rim of coated pits and that Cbl-induced ubiquitination of the EGFR allows relocation of EGFR from the rim to the center of clathrin-coated pits.

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Section: Discussionmentioning

confidence: 77%

Cbl-dependent Ubiquitination Is Required for Progression of EGF Receptors into Clathrin-coated Pits

Stang

Blystad

Kazazic

et al. 2004

MBoC

146

168

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“…Unlike the amphiphysins and endophilins, Rvs167 does not carry clathrin and clathrin adaptor (AP)-binding motifs and does not appear to bind either of these proteins. This difference may exist because clathrin adaptors are not required for receptor internalization in yeast (61,62), and clathrin itself is not strictly required (63).…”

Section: Discussionmentioning

confidence: 99%

The Rsp5 Ubiquitin Ligase Binds to and Ubiquitinates Members of the Yeast CIN85-Endophilin Complex, Sla1-Rvs167

Stamenova¹,

Dunn²,

Adler

et al. 2004

Journal of Biological Chemistry

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Sla1 and Rvs167 are yeast proteins required for receptor internalization and organization of the actin cytoskeleton. Here we provide evidence that Sla1 and Rvs167 are orthologues of the mammalian CIN85 and endophilin proteins, respectively, which are required for ligand-stimulated growth factor receptor internalization. Sla1 is similar in domain structure to CIN85 and binds directly to the endophilin-like Rvs167. Akin to CIN85, Sla1 interacts with synaptojanins and a ubiquitin ligase that regulates endocytosis. This ubiquitin ligase, Rsp5, binds directly to both Sla1 and Rvs167. The interaction between Rsp5 and Rvs167 is mediated through Rsp5 WW domains and PXY motifs in the central Gly-Pro-Ala-rich domain of Rvs167. Rvs167 PXY motifs are required for Rsp5-dependent monoubiquitination of Rvs167 on Lys 481 in the Src homology 3 (SH3) domain. Mutation of Lys 481 3 Arg causes cells to grow slowly on medium containing 1 M NaCl, although this phenotype is not due to the defect in ubiquitination caused by the K481R mutation. We propose that Rsp5 interaction with Sla1-Rvs167 promotes Rvs167 ubiquitination and regulates activity of this protein complex. Rvs167 ubiquitination is not required for general function of Rvs167, but may control specific Rvs167 SH3 domain-protein interactions or negatively regulate SH3 domain activity.

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“…For example, the type I-like sequence 451 LIDL(COOH) in the Saccharomyces cerevisiae epsin homologues Ent1p and Ent2p (52) could explain the unexpected presence of clathrin-coated vesicles in strains engineered to lack functional forms of the yeast heterotetrameric adaptors and AP180 (53,54). In mammalian cells, overexpression of an AP-2-binding region from amphiphysin I grossly disrupts the intracellular localization of AP-2 but, surprisingly, appears to leave the normal clathrin distribution at the cell surface intact (16).…”

Section: Fig 3 Direct Interaction Of Adaptors With Gst-pwdlwmentioning

confidence: 99%

Interaction of Two Structurally Distinct Sequence Types with the Clathrin Terminal Domain β-Propeller

Drake¹,

Traub²

2001

Journal of Biological Chemistry

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The amino-terminal domain of the clathrin heavy chain, which folds into a seven-bladed ␤-propeller, binds directly to several endocytic proteins via short sequences based on the consensus residues LLDLD. In addition to a single LLDLD-based, type I clathrin-binding sequence, both amphiphysin and epsin contain a second, distinct sequence that is also capable of binding to clathrin directly. Here, we analyzed these sequences, which we term type II sequences, and show that the 257 LMDLA sequence in rat epsin 1 appears to be a weak clathrin-binding variant of the sequence 417 PWDLW originally found in human amphiphysin II. The structural features of the type II sequence required for association with clathrin are distinct from the LLDLD-based sequence. In the central segment of amphiphysin, the type I and type II sequences cooperate to effect optimal clathrin binding and the formation of sedimentable assemblies. Together, the data provide evidence for two interaction surfaces upon certain endocytic accessory proteins that could cooperate with other coat components to enhance clathrin bud formation at the cell surface.The most characteristic feature of assembled clathrin is the polygonal appearance of the coat. This largely reflects the geometric arrangement of the clathrin molecule. During biosynthesis, three ϳ190-kDa clathrin heavy chains trimerize via the carboxyl-terminal end to form a characteristic triskelion. An ϳ25-kDa light chain also binds to each heavy chain, near the site of trimerization, the central vertex. Each heavy chain projects out radially from the vertex, with the three legs of the trimer splayed ϳ120°apart. The triskelion leg is a relatively rigid structure, composed of tandemly stacked repeats of ␣ zigzags (1, 2). Lateral packing of the adjacent helix pairs generates an extended linear rod, interrupted by a kink roughly halfway along the length (3). Clathrin trimers also display an inherent sidedness (4). When viewed from above, the kink redirects the distal segment of each leg clockwise and down, positioning the globular ϳ350-residue amino-terminal portion, the terminal domain, of each heavy chain below the plane of both the vertex and the proximal portion of each leg (5).As clathrin trimers begin to associate to form a coat, packing occurs by the antiparallel apposition of the proximal portion of one triskelion leg with the proximal segment of an adjacent leg. The distal segments of two other assembling trimers pack below the antiparallel proximal leg pair (6). This leads to the generation of the hexagonal and pentagonal facets that typify the clathrin coat. Truncated trimers termed clathrin hubs, composed of roughly the carboxyl-terminal third of the heavy chain, can also assemble into pseudolattices at reduced pH in the presence of calcium (7). These assemblies do not display the characteristic curvature seen in clathrin coats, however, and do not form closed spherical structures (7). Because heavy chainheavy chain leg interactions are relatively weak at physiological pH, these observations p...

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Clathrin functions in the absence of heterotetrameric adaptors and AP180-related proteins in yeast

Cited by 133 publications

References 100 publications

Cbl-dependent Ubiquitination Is Required for Progression of EGF Receptors into Clathrin-coated Pits

Cbl-dependent Ubiquitination Is Required for Progression of EGF Receptors into Clathrin-coated Pits

The Rsp5 Ubiquitin Ligase Binds to and Ubiquitinates Members of the Yeast CIN85-Endophilin Complex, Sla1-Rvs167

Interaction of Two Structurally Distinct Sequence Types with the Clathrin Terminal Domain β-Propeller

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