Coatomer Interaction with Di-Lysine Endoplasmic Reticulum Retention Motifs

Cosson, Pierre; Letourneur, François

doi:10.1126/science.8128252

Cited by 548 publications

(462 citation statements)

References 23 publications

Supporting

Mentioning

444

Contrasting

Order By: Relevance

“…GST fusions in which the lysines were mutated to serines were used as controls. These experiments used whole cell extracts from cells grown at 24°C, and the high salt/0.5% Triton X-100 binding conditions described by Cosson and Letourneur (1994). We analyzed extracts from the mutants sec27 -95, sec27⌬1-285, ret1-1, and ret1⌬1-285, and from wild-type cells.…”

Section: Binding Of Coatomer From ␣-And ␤ -Cop Mutants To Di-lysine Mmentioning

confidence: 99%

“…This di-lysine motif is necessary and sufficient to target reporter proteins into the retrograde pathway in vivo, and coatomer can directly interact with such motifs in vitro (Jackson et al, 1993;Gaynor et al, 1994;Cosson and Letourneur, 1994;Schrö der-Kohne et al, 1998). Using reporter proteins tagged with variants of di-lysine motifs, it has been previously demonstrated clearly that small changes in the local sequence context of the lysines can dramatically alter the signal strength of the di-lysine motif and thus generate a broad spectrum of trafficking phenotypes (Zerangue et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The α- and β′-COP WD40 Domains Mediate Cargo-selective Interactions with Distinct Di-lysine Motifs

Eugster

Frigerio

Dale

et al. 2004

MBoC

102

107

View full text Add to dashboard Cite

Coatomer is required for the retrieval of proteins from an early Golgi compartment back to the endoplasmic reticulum. The WD40 domain of ␣-COP is required for the recruitment of KKTN-tagged proteins into coatomer-coated vesicles. However, lack of the domain has only minor effects on growth in yeast. Here, we show that the WD40 domain of ␤ -COP is required for the recycling of the KTKLL-tagged Golgi protein Emp47p. The protein is degraded more rapidly in cells with a point mutation in the WD40 domain of ␤ -COP (sec27-95) or in cells lacking the domain altogether, whereas a point mutation in the Clathrin Heavy Chain Repeat (sec27-1) does not affect the turnover of Emp47p. Lack of the WD40 domain of ␤ -COP has only minor effects on growth of yeast cells; however, absence of both WD40 domains of ␣-and ␤ -COP is lethal. Two hybrid studies together with our analysis of the maturation of KKTN-tagged invertase and the turnover of Emp47p in ␣-and ␤ -COP mutants suggest that the two WD40 domains of ␣-and ␤ -COP bind distinct but overlapping sets of di-lysine signals and hence both contribute to recycling of proteins with di-lysine signals.

show abstract

Section: Binding Of Coatomer From ␣-And ␤ -Cop Mutants To Di-lysine Mmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The α- and β′-COP WD40 Domains Mediate Cargo-selective Interactions with Distinct Di-lysine Motifs

Eugster

Frigerio

Dale

et al. 2004

MBoC

102

107

View full text Add to dashboard Cite

show abstract

“…GST-pull down experiments were carried out as described (Cosson and Letourneur, 1994), but yeast cells were lysed in PBS, 10 mM EDTA, 0.5% Triton, protease inhibitor cocktail, and PMSF. For binding to ubiquitin-sepharose, 50 l ubiquitin-sepharose (Sigma) was incubated with 20 g recombinant protein for 4 h, and washed in TBST as described (Aguilar et al, 2003).…”

Section: Pull-down Experiments and Ubiquitin-binding Assaysmentioning

confidence: 99%

Ent5p Is Required with Ent3p and Vps27p for Ubiquitin-dependent Protein Sorting into the Multivesicular Body

Eugster¹,

Pécheur²,

Michel

et al. 2004

MBoC

Self Cite

View full text Add to dashboard Cite

At the late endosomes, cargoes destined for the interior of the vacuole are sorted into invaginating vesicles of the multivesicular body. Both PtdIns(3,5)P 2 and ubiquitin are necessary for proper sorting of some of these cargoes. We show that Ent5p, a yeast protein of the epsin family homologous to Ent3p, localizes to endosomes and specifically binds to PtdIns(3,5)P 2 via its ENTH domain. In cells lacking Ent3p and Ent5p, ubiquitin-dependent sorting of biosynthetic and endocytic cargo into the multivesicular body is disrupted, whereas other trafficking routes to the vacuole are not affected. Ent3p and Ent5p are associated with Vps27p, a FYVE domain containing protein that interacts with ubiquitinated cargoes and is required for protein sorting into the multivesicular body. Therefore, Ent3p and Ent5p are the first proteins shown to be connectors between PtdIns(3,5)P 2 -and the Vps27p-ubiquitin-driven sorting machinery at the multivesicular body. INTRODUCTIONEndosomes are crossroads between the biosynthetic and the endocytic pathways. Here, proteins destined for the lysosome/vacuole are segregated away from proteins recycling back to the plasma membrane or to the Golgi apparatus. A critical membrane segregation and protein-sorting event takes place in late endosomes, regions of endosomal membranes invaginate and vesicles bud into the lumen of the organelle, giving rise to the multivesicular body (MVB;Felder et al., 1990). Mature MVB fuses with the vacuole/ lysosome releasing internal vesicles into its lumen and thereby exposing them to the activity of hydrolytic enzymes (Futter et al., 1996;Odorizzi et al., 1998). This mechanism allows a subset of endosomal membrane proteins to be sorted to the lumen of the lysosome/vacuole, whereas others are either selectively recycled back to the Golgi or to the plasma membrane, or they end up on the lysosomal/vacuolar membrane (Pelham, 2002). In yeast, the MVB pathway is crucial for the regulated turnover of pheromone receptors and of some permeases (Odorizzi et al., 1998). Further, biosynthetic transmembrane proteins such as carboxypeptidase S (Cps1p) and Phm5p follow this route to reach their final destination, the vacuolar lumen (Odorizzi et al., 1998;Reggiori and Pelham, 2001;Epple et al., 2003).Ubiquitination has been implicated as an endosomal-sorting signal. Cps1p and Phm5p ubiquitination is essential to target them into the vacuolar lumen after sorting at the MVB (Katzmann et al., 2001;Reggiori and Pelham, 2001). Potential cargo receptors responsible for recognition and recruitment of ubiquitinated cargo into MVB vesicles are class E Vps (vacuolar protein sorting) proteins. Class E mutants accumulate cargo destined for the vacuole in an exaggerated endosomal structure and all (currently identified) class E proteins are required for sorting at the MVB (Conibear and Stevens, 1998). Vps23p, a class E protein of the ESCRT-I complex (endosomal-sorting complex required for transport), carries an ubiquitin-conjugating (UBC)-like domain and interacts directly with ubiquitin i...

show abstract

“…Both proteins have a type I transmembrane topology, and a stretch of about 200 amino acids in their luminal domain shows 19-24% identity to leguminous lectins. Their short cytoplasmic domains contain related sorting signals that are likely to interact with cytoplasmic coats, either coatomer or clathrin adaptors (Cosson and Letourneur, 1994;Letourneur et al, 1994;Itin et al, 1995a,b). The two proteins are differently distributed in the endomembrane system.…”

Section: Introductionmentioning

confidence: 99%

ERGIC-53 is a functional mannose-selective and calcium-dependent human homologue of leguminous lectins.

Itin

Roche

Monsigny

et al. 1996

MBoC

166

148

View full text Add to dashboard Cite

Based on sequence homologies with leguminous lectins, the intermediate compartment marker proposed to be a member of a putative new class of animal lectins associated with the secretory pathway. Independently, a promyelocytic protein, MR60, was purified by mannose-column chromatography, and a cDNA was isolated that matched MR60 peptide sequences. This cDNA was identical to that of and homologies with the animal lectin family of the galectins were noticed. Not all peptide sequences of MR60, however, were found in ERGIC-53, raising the possibility that another protein associated with ERGIC-53 may possess the lectin activity. Here, we provide the first direct evidence for a lectin function of ERGIC-53. Overexpressed ERGIC-53 binds to a mannose column in a calcium-dependent manner and also co-stains with mannosylated neoglycoprotein in a morphological binding assay. By using a sequential elution protocol we show that ERGIC-53 has selectivity for mannose and low affinity for glucose and GlcNAc, but no affinity for galactose. To experimentally address the putative homology of ERGIC-53 to leguminous lectins, a highly conserved protein family with an invariant asparagine essential for carbohydrate binding, we substituted the corresponding asparagine in ERGIC-53. This mutation, as well as a mutation affecting a second site in the putative carbohydrate recognition domain, abolished mannosecolumn binding and co-staining with mannosylated neoglycoprotein. These findings establish ERGIC-53 as a lectin and provide functional evidence for its relationship to leguminous lectins. Based on its monosaccharide specificity, domain organization, and recycling properties, we propose ERGIC-53 to function as a sorting receptor for glycoproteins in the early secretory pathway.

show abstract

Coatomer Interaction with Di-Lysine Endoplasmic Reticulum Retention Motifs

Cited by 548 publications

References 23 publications

The α- and β′-COP WD40 Domains Mediate Cargo-selective Interactions with Distinct Di-lysine Motifs

The α- and β′-COP WD40 Domains Mediate Cargo-selective Interactions with Distinct Di-lysine Motifs

Ent5p Is Required with Ent3p and Vps27p for Ubiquitin-dependent Protein Sorting into the Multivesicular Body

ERGIC-53 is a functional mannose-selective and calcium-dependent human homologue of leguminous lectins.

Contact Info

Product

Resources

About