A tyrosine-containing motif mediates ER retention of CD3-epsilon and adopts a helix-turn structure.

Mallabiabarrena, Arrate; Jiménez, M.; Rico, Manuel; Alarcón, Balbino

doi:10.1002/j.1460-2075.1995.tb07220.x

Cited by 64 publications

(61 citation statements)

References 61 publications

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“…While there are well-known examples of retrieval signals (Nilsson & Warren, 1994;Nilsson et al, 1989;Pelham, 1988), only a few true (static) retention motifs have been identified (Cocquerel et al, 1998(Cocquerel et al, , 1999. The fact that NSP4 does not contain the established ER retrieval-retention signal for type 1 transmembrane proteins -two lysines at 23 and 24 or 25 position (Jackson et al, 1993) or a tyrosinebased signal motif (Mallabiabarrena et al, 1995) -could indicate that NSP4 may contain a motif for a 'static retention signal' or an as yet unknown retrieval signal. Taylor et al (1996) have shown that cytoplasmic tail of NSP4 not only binds to microtubules but also inhibits anterograde protein trafficking, as illustrated by the inhibition of transport of VSV-G protein to the plasma membrane (Xu et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…Three sorting signals associated with ER retention and retrieval (Jackson et al, 1993;Lewis & Pelham, 1992;Pelham, 1994;Townsley & Pelham, 1994) have been proposed: (i) soluble proteins in the ER lumen carry a specific C terminus tetrapeptide retention signal, H/KDEL (Munro & Pelham, 1987); (ii) transmembrane proteins (type II) carry two arginines near the cytoplasmic N terminus (Schutze et al, 1994); and (iii) transmembrane proteins (type I) possess lysines at positions 23 and either 24 or 25 (Jackson et al, 1990). A tyrosine-based motif close to the C terminus has also been identified as an ER retention signal (Mallabiabarrena et al, 1995), but the retention mechanism for this motif is not yet known. In addition to such retrieval mechanisms, retention in the ER may also be static for glycoproteins that do not contain any of the above signal motifs (Cocquerel et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

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A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum

Mirazimi

Magnusson

Svensson

2003

Journal of General Virology

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The rotavirus genome encodes two glycoproteins, one structural (VP7) and one non-structural (NSP4), both of which mature and remain in the endoplasmic reticulum (ER). While three amino acids in the N terminus have been proposed to function as a retention signal for VP7, no information is yet available on how NSP4 remains associated with the ER. In this study, we have investigated the ER retention motif of NSP4 by producing various C-terminal truncations. Deleting the C terminus by 52 amino acids did not change the intracellular distribution of NSP4, but an additional deletion of 38 amino acids diminished the ER retention and resulted in the expression of NSP4 on the cell surface. Brefeldin A treatment prevented NSP4 from reaching the cell surface, suggesting that C-terminal truncated plasma membrane NSP4 is transported through the normal secretory pathway. On the basis of these results, we propose that the region between amino acids 85 and 123 in the cytoplasmic region of NSP4 are involved in ER retention. INTRODUCTIONThe secretory pathway of eukaryotic cells provides a route for proteins to be secreted from the cells and to be targeted to various cellular organelles (Jackson et al., 1990;Nilsson & Warren, 1994;Paabo et al., 1987). Following synthesis on the endoplasmic reticulum (ER), certain proteins resist the bulk flow of the secretory pathway and become ER-associated and accumulate in the ER. Three sorting signals associated with ER retention and retrieval (Jackson et al., 1993;Lewis & Pelham, 1992;Pelham, 1994;Townsley & Pelham, 1994) have been proposed: (i) soluble proteins in the ER lumen carry a specific C terminus tetrapeptide retention signal, H/KDEL (Munro & Pelham, 1987); (ii) transmembrane proteins (type II) carry two arginines near the cytoplasmic N terminus (Schutze et al., 1994); and (iii) transmembrane proteins (type I) possess lysines at positions 23 and either 24 or 25 (Jackson et al., 1990). A tyrosine-based motif close to the C terminus has also been identified as an ER retention signal (Mallabiabarrena et al., 1995), but the retention mechanism for this motif is not yet known. In addition to such retrieval mechanisms, retention in the ER may also be static for glycoproteins that do not contain any of the above signal motifs (Cocquerel et al., 1999).Rotavirus undergoes a unique maturation process in the ER. The assembly process, which includes translocation of doublelayered particles across the ER and retention of mature virus in the ER, has provided a system in which post-translational events can be studied (Bellamy & Both, 1990;Estes & Cohen, 1989;Poruchynsky et al., 1985Poruchynsky et al., , 1991. Two rotavirus proteins have been in focus with regard to assembly and pathogenesis, the VP7 outer capsid protein, which is luminal and ERassociated, and NSP4, a non-structural glycoprotein that has been given significant attention in recent years. NSP4 is a novel type of trans-ER resident glycoprotein, which functions not only as a receptor for double-layered particles in the cytoplasm (Au et a...

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum

Mirazimi

Magnusson

Svensson

2003

Journal of General Virology

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“…The T-cell receptor delta chain displays a GYYYYV sequence in its membranous segment (33). That a tyrosine-containing motif mediates ER retention of CD3-⑀ chain has also been reported (34). To determine the topogenic importance of the tyrosyl residue cluster GFP chimera, Construct d lacking the tyrosyl residues in region II was generated.…”

Section: Fig 3 Electron Microscopic Immunogold Labeling Of Rab E3 Nmentioning

confidence: 99%

Targeting Proteins to the Lumen of Endoplasmic Reticulum Using N-terminal Domains of 11β-Hydroxysteroid Dehydrogenase and the 50-kDa Esterase

Mziaut¹,

Korza²,

Hand³

et al. 1999

Journal of Biological Chemistry

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Previous studies identified two intrinsic endoplasmic reticulum (ER) proteins, 11␤-hydroxysteroid dehydrogenase, isozyme 1 (11␤-HSD) and the 50-kDa esterase (E3), sharing some amino acid sequence motifs in their N-terminal transmembrane (TM) domains. Both are type II membrane proteins with the C terminus projecting into the lumen of the ER. This finding implied that the N-terminal TM domains of 11␤-HSD and E3 may constitute a lumenal targeting signal (LTS). To investigate this hypothesis we created chimeric fusions using the putative targeting sequences and the reporter gene, Aequorea victoria green fluorescent protein. Transfected COS cells expressing LTS-green fluorescent protein chimeras were examined by fluorescent microscopy and electron microscopic immunogold labeling. The orientation of expressed chimeras was established by immunocytofluorescent staining of selectively permeabilized COS cells. In addition, protease protection assays of membranes in the presence and absence of detergents was used to confirm lumenal or the cytosolic orientation of the constructed chimeras. To investigate the general applicability of the proposed LTS, we fused the N terminus of E3 to the N terminus of the NADH-cytochrome b5 reductase lacking the myristoyl group and N-terminal 30-residue membrane anchor. The orientation of the cytochrome b5 reductase was reversed, from cytosolic to lumenal projection of the active domain. These observations establish that an amino acid sequence consisting of short basic or neutral residues at the N terminus, followed by a specific array of hydrophobic residues terminating with acidic residues, is sufficient for lumenal targeting of single-pass proteins that are structurally and functionally unrelated.

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“…As a control, a truncated mutant of CD8␣ lacking the cytoplasmic tail was used (CD8t). Although the CD8/⑀ construct is in part retained in the ER because the cytoplasmic tail of CD3⑀ contains an ER retention sequence (14,15), both CD8/⑀ and CD8t were expressed on the cell surface. This allowed separation of transfected COS cells from untransfected cells by immunoselection with antibody-coated magnetic beads.…”

Section: Expression Of a Protein Chimera Containing The Cytoplasmic Tmentioning

confidence: 99%

“…The proline-rich sequence contains the SH3-binding consensus motif XPPXP, and the C-terminal region contains the YXXLXXR endoplasmic reticulum (ER) retention sequence, which partially overlaps the ITAM (14,15). Previous attempts to identify proteins that associate with the cytoplasmic tail of CD3⑀ have shown the specific interaction of a nuclear protein, topoisomerase II␤, and a tyrosine-phosphorylated protein, CAST, with the N-terminal region of the CD3⑀ tail (13,16).…”

mentioning

confidence: 99%

Intracellular Redistribution of Nucleolin upon Interaction with the CD3ε Chain of the T Cell Receptor Complex

Gil

Gutiérrez

Alarcón

2001

Journal of Biological Chemistry

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T cell activation through the antigen receptor (TCR) involves the cytoplasmic tails of the CD3 subunits CD3␥, CD3␦, CD3⑀, and CD3. Whereas the biological significance of the cytoplasmic tails of these molecules is suggested, in part, by their evolutionarily conserved sequences, their interactions with signal transduction molecules are not completely understood. We used affinity chromatography columns of glutathione S-transferase fused to the CD3⑀ cytoplasmic tail to isolate proteins that specifically interact with this subunit. In this way, we identified the shuttling protein nucleolin as a specific CD3⑀-interacting molecule. Using competition studies and affinity chromatography on peptide columns, we were able to identify a central proline-rich sequence as the nucleolin-interacting sequence in CD3⑀. Transfection in COS cells of wild type CD3⑀, but not of nonbinding mutants of CD3⑀, resulted in redistribution of nucleolin from the nucleus and nucleoli to the cytoplasm. This property was transferred to a CD8 protein chimera by appending the cytoplasmic tail of CD3⑀. We also found that nucleolin associated with the TCR complex. This association was increased upon TCR engagement, suggesting that the CD3⑀/nucleolin interaction may have a role in T cell activation.T cells respond to antigen via a polypeptide complex composed of ligand-binding T cell receptor (TCR) 1 ␣ and ␤ chains (or ␥ and ␦ in ␥␦ T cells) and the CD3 subunits CD3␥, CD3␦, CD3⑀, and CD3 (1, 2). Unlike the TCR chains, the CD3 components have long cytoplasmic tails that associate with cytoplasmic signal transduction molecules. This association is mediated at least in part by a double tyrosine-based motif present in a single copy in the CD3␥, CD3␦, and CD3⑀ chains and in three copies in CD3 (3). This motif, named immune-receptor tyrosine-based activation motif (ITAM), becomes tyrosine phosphorylated during T cell activation by the Src family proteintyrosine kinases . Tyrosine phosphorylated ITAM become docking sites for the Syk family proteintyrosine kinase ZAP70 and other signal-transducing molecules. It is well established that antibody-mediated engagement of protein chimeras containing the cytoplasmic tail of either CD3 or CD3⑀ results in T cell activation (7-10). These data indicate that the cytoplasmic tail of one of these subunits can be sufficient to induce T cell activation. Regarding the role of CD3 subunits in T cell activation, most of the attention has been focused on the ITAM. However, the cytoplasmic tails of the CD3 subunits contain other evolutionarily conserved features that suggest ITAM-independent roles for them.The CD3⑀ cytoplasmic tail, highly conserved (11, 12), can be tentatively subdivided into three regions; the N-terminal region contains a basic amino acid cluster, the central region contains a proline-rich sequence, and the C-terminal region contains the ITAM (13). The proline-rich sequence contains the SH3-binding consensus motif XPPXP, and the C-terminal region contains the YXXLXXR endoplasmic reticulum (ER) retention sequ...

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A tyrosine-containing motif mediates ER retention of CD3-epsilon and adopts a helix-turn structure.

Cited by 64 publications

References 61 publications

A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum

A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum

Targeting Proteins to the Lumen of Endoplasmic Reticulum Using N-terminal Domains of 11β-Hydroxysteroid Dehydrogenase and the 50-kDa Esterase

Intracellular Redistribution of Nucleolin upon Interaction with the CD3ε Chain of the T Cell Receptor Complex

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