Different Fate of a Single Reporter Protein Containing KDEL or KKXX Targeting Signals Stably Expressed in Mammalian Cells

Martire, Gianluca; Mottola, Giovanna; Pascale, Maria; Malagolini, Nadia; Turrini, Ileana; Serafini‐Cessi, Franca; Jackson, Michael R.; Bonatti, Stefano

doi:10.1074/jbc.271.7.3541

Cited by 37 publications

(41 citation statements)

References 52 publications

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“…Although CD4-TM1E and CD4-TM1prM were localized in the ER at steady state, our data do not exclude transport to the cis Golgi followed by recycling to the ER, as observed for luminal proteins containing a C-terminal sequence of the prototype KDEL (Lewis & Pelham, 1992;Martire et al, 1996;Townsley et al, 1993). To answer this question, cells expressing CD4-TM1E or CD4-TM1prM were treated with nocodazole.…”

Section: Localization Signals In Tm1 Of E and Prm Induce A Static Retmentioning

confidence: 95%

Identification of a dominant endoplasmic reticulum-retention signal in yellow fever virus pre-membrane protein

Ciczora

Callens

Séron

et al. 2009

Journal of General Virology

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Yellow fever virus (YFV) encodes two envelope proteins, pre-membrane (prM) and envelope (E), that accumulate in the endoplasmic reticulum (ER). The C termini of prM and E form two antiparallel transmembrane a-helices that contain ER-retention signals. To understand further the ER retention of the prME heterodimer, we characterized the subcellular localization of chimeric proteins made of a reporter protein fused to the transmembrane segments of YFV envelope proteins. We showed that at least three of the transmembrane segments of the prME heterodimer are ER-retention signals. Interestingly, increasing the length of these a-helices led to the export of the chimeric proteins out of the ER. Furthermore, adding a diacidic export signal at the C terminus of the first transmembrane segment of the E protein also induced export to the cell surface. However, adding this export signal at the C terminus of the first transmembrane segment of E in the context of prME did not change the subcellular localization of the prME heterodimer, suggesting the presence of a stronger ER-retention signal outside the first transmembrane segment of E. Importantly, the diacidic export motif added to the C terminus of the first transmembrane segment of the prM protein was not sufficient to export a chimeric protein out of the ER, indicating that this sequence is a dominant ER-retention signal. Together, these data indicate that a combination of several signals of different strengths contributes to the ER retention of the YFV envelope protein heterodimer.

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Section: Localization Signals In Tm1 Of E and Prm Induce A Static Retmentioning

confidence: 95%

Identification of a dominant endoplasmic reticulum-retention signal in yellow fever virus pre-membrane protein

Ciczora

Callens

Séron

et al. 2009

Journal of General Virology

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“…This motif has been identified to be a retrieval motif that brings proteins from an unknown sorting compartment back to the ER (Jackson et al, 1990). It is shown that the KKXX-mediated retrieval can occur at several sites between the intermediate compartment and the trans-Golgi network (Jackson et al, 1993;Martire et al, 1996). COOHterminal dilysine-motifs bind specifically to the protein complex coatomer Letourneur et al, 1994;Lowe and Kreis, 1995), which is essential for the retrieval of dilysine-tagged proteins to the ER.…”

Section: Discussionmentioning

confidence: 99%

Tmp21 and p24A, Two Type I Proteins Enriched in Pancreatic Microsomal Membranes, Are Members of a Protein Family Involved in Vesicular Trafficking

Blum

Feick

Puype

et al. 1996

Journal of Biological Chemistry

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We report here on the isolation, cloning, and expression of two M r 21,000 proteins from rat pancreatic acinar cells, the rat-Tmp21 (transmembrane protein, M r 21,000) and the rat-p24A. Both proteins are transmembrane proteins with type I topology and share weak but significant homology to one another (23% identity). We further show the cloning and characterization of the human homologs, hum-Tmp21, which is expressed in two variants (Tmp21-I and Tmp21-II), and hum-p24A. Tmp21 proteins and p24A have highly conserved COOH-terminal tails, which contain motifs related to the endoplasmic reticulum retention and retrieval consensus sequence KKXX. The rat-p24 sequence is identical to the hamster

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“…Antibodies-The following antibodies were used: the OKT8 mouse anti-CD8 monoclonal antibody, from Ortho (Raritan, NJ), the N1 mouse anti-CD8 monoclonal antibody (29), a rabbit anti-ERGIC-53 antibody (30), a rabbit anti-GM130 antibody, a rabbit anti-Sec31 antibody, and a rabbit anti-giantin antibody (31), a mouse anti-hemagglutinin antibody, and a rabbit antihemagglutinin antibody from Santa Cruz. The anti-GRASP65 polyclonal antibody was obtained through immunization against a fusion protein between glutathione S-transferase (GST) and rat GRASP65 and affinity purified; the anti-GRASP55 polyclonal antibody was a kind gift of Dr. F. Barr.…”

Section: Methodsmentioning

confidence: 99%

GRASP65 and GRASP55 Sequentially Promote the Transport of C-terminal Valine-bearing Cargos to and through the Golgi Complex

D’Angelo

Prencipe

Iodice

et al. 2009

Journal of Biological Chemistry

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The Golgi matrix proteins GRASP65 and GRASP55 have recognized roles in maintaining the architecture of the Golgi complex, in mitotic progression and in unconventional protein secretion whereas, surprisingly, they have been shown to be dispensable for the transport of commonly used reporter cargo proteins along the secretory pathway. However, it is becoming increasingly clear that many trafficking machineries operate in a cargo-specific manner, thus we have investigated whether GRASPs may control the trafficking of selected classes of cargo. We have taken into consideration the C-terminal valine-bearing receptors CD8␣ and Frizzled4 that we show bind directly to the PSD95-DlgA-zo-1 (PDZ) domains of GRASP65 and GRASP55. We demonstrate that both GRASPs are needed sequentially for the efficient transport to and through the Golgi complex of these receptors, thus highlighting a novel role for the GRASPs in membrane trafficking. Our results open new perspectives for our understanding of the regulation of surface expression of a class of membrane proteins, and suggests the causal mechanisms of a dominant form of autosomal human familial exudative vitreoretinopathy that arises from the Frizzled4 mutation involving its C-terminal valine.GRASP65 and GRASP55 were identified in in vitro assays as factors that are required for the stacking of the Golgi cisternae (1, 2). This activity arose as the result of the tethering functions displayed by GRASP65 and GRASP55, through their interactions with their partner proteins GM130 and golgin-45, respectively (2-4). Several other studies have shown more recently that the GRASPs are involved in the maintenance of the structure of the Golgi ribbon in mammal cells during interphase, in controlling the fragmentation of the Golgi complex at the onset of mitosis (5-8), in establishing cell polarity in migrating cells (9), and in the consumption of COPII vesicles and the formation of the cis-Golgi in yeast (10).Although the role of the GRASPs in control of the Golgi complex structure (both in interphase and during mitosis) is well established, their involvement in cargo transport along the secretory pathway is still debated. Indeed, a direct role for GRASPs in cargo transport has so far been established only for the unconventional secretion routes in Dictyostilium and Drosophila (11, 12), whereas they have been shown not to be directly involved in the trafficking of commonly studied reporter cargo proteins along the "conventional" secretory pathway (e.g. the temperature-sensitive (ts-045) mutant of the G protein of vesicular stomatitis virus (VSVG) 2 ) and secretory horseradish peroxidase (5, 6, 13, 14). GRASPs can engage different types of interactions including the ones mediated by their PDZ domains, through which the GRASPs cannot only homodimerize, thus participating in cisternal stacking (15) but can also bind the C-terminal valine motifs (C-TVM) of membrane proteins such as protransforming growth factor ␣ and p24a (16,17).Interestingly a C-terminal valine can behave as "transport sign...

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Different Fate of a Single Reporter Protein Containing KDEL or KKXX Targeting Signals Stably Expressed in Mammalian Cells

Cited by 37 publications

References 52 publications

Identification of a dominant endoplasmic reticulum-retention signal in yellow fever virus pre-membrane protein

Identification of a dominant endoplasmic reticulum-retention signal in yellow fever virus pre-membrane protein

Tmp21 and p24A, Two Type I Proteins Enriched in Pancreatic Microsomal Membranes, Are Members of a Protein Family Involved in Vesicular Trafficking

GRASP65 and GRASP55 Sequentially Promote the Transport of C-terminal Valine-bearing Cargos to and through the Golgi Complex

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