Role of Sec24 isoforms in selective export of membrane proteins from the endoplasmic reticulum

Wendeler, Markus W.; Paccaud, Jean‐Pierre; Hauri, Hans‐Peter

doi:10.1038/sj.embor.7400893

Cited by 168 publications

(175 citation statements)

References 19 publications

(23 reference statements)

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“…Four Sec24 isoforms, designated Sec24A-D, are expressed in mammalian systems (54) and proteins with close sequence matches to all four human Sec24 proteins are present in the chicken database. Human Sec24 isoforms show different but overlapping cargo binding preferences, and Sec24A is most important for ER export (55). It is possible that a functional difference between Sec24 proteins exists in DT40, and Sec23 is more greatly associated with another isoform rather than the Sec24C homolog we identified.…”

Section: Resultsmentioning

confidence: 83%

The Organelle Proteome of the DT40 Lymphocyte Cell Line

Hall

Hester

Griffin

et al. 2009

Molecular & Cellular Proteomics

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A major challenge in eukaryotic cell biology is to understand the roles of individual proteins and the subcellular compartments in which they reside. Here, we use the localization of organelle proteins by isotope tagging technique to complete the first proteomic analysis of the major organelles of the DT40 lymphocyte cell line. This cell line is emerging as an important research tool because of the ease with which gene knockouts can be generated. We identify 1090 proteins through the analysis of preparations enriched for integral membrane or soluble and peripherally associated proteins and localize 223 proteins to the endoplasmic reticulum, Golgi, lysosome, mitochondrion, or plasma membrane by matching their density gradient distributions to those of known organelle residents. A striking finding is that within the secretory and The chicken pre-B cell line DT40 exhibits a remarkably high ratio of targeted to random integration for transfected DNA constructs. This property is unusual in vertebrate cell lines and enables targeted gene disruption experiments to be carried out with relative ease (1). Consequently, DT40 has become a major research tool for the molecular dissection of a wide range of cellular and biochemical mechanisms in a vertebrate context, including membrane traffic, signal transduction, and cell cycle (2).Proteins in eukaryotic cells are organized according to their functions within a dynamic network of membranes. Localization is therefore paramount in assigning functions to uncharacterized proteins and understanding the processes occurring in subcellular compartments. An increased knowledge of the protein localization within the DT40 cell line would be of great value. Traditional localization methods such as immunofluorescence microscopy are typically low throughput and are more suitably applied to the study of specific proteins of interest rather than the cataloguing of large numbers of proteins. Recent developments in proteomics have made it possible to analyze the protein composition of organelles using a variety of different approaches. Several groups have utilized label-free quantitative proteomics in the high throughput assignment of proteins to subcellular compartments. In one approach, protein correlation profiling, proteins from enriched organelle fractions are quantified by peptide ion intensity measurements (3, 4). Other similar methods employ quantitation by spectral counting, recording the number of ions detected per protein (5, 6). Localization of organelle proteins by isotope tagging (LOPIT) 1 is a complementary approach, which employs isotope labeling for quantitation (7-9). Rather than processing each sample separately as in label-free techniques, differentially labeled fractions are pooled early in the LOPIT protocol. This has the important advantage of reducing the points at which variation might be introduced into the data.LOPIT begins with the partial separation of organelles by density gradient centrifugation and relies on the assumption that proteins from each organelle c...

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Section: Resultsmentioning

confidence: 83%

The Organelle Proteome of the DT40 Lymphocyte Cell Line

Hall

Hester

Griffin

et al. 2009

Molecular & Cellular Proteomics

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“…In yeast cells, the deletion of LST1, encoding an isoform of Sec24, inhibits the trafficking of Pma1p from the ER but does not affect the trafficking of other proteins (Roberg et al, 1999). In animal cells, Wendeler et al (2007) reported the differential effects of Sec24 isoform-specific silencing on the transport of membrane proteins. Similarly, in Arabidopsis, multiple isoforms of Sec24 exist, supporting the possibility that multiple types of COPII vesicles also exist in plant cells.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, one possible explanation for the differential inhibition is that plant cells may contain multiple types of COPII vesicles that are differentially used depending on the cargo proteins. Indeed, in animal cells and yeast cells, multiple types of COPII vesicles exist (Tang et al, 1999;Wendeler et al, 2007). Among COPII vesicle components, Sec24 plays a critical role in selecting cargo proteins for the COPII vesicle (Miller et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

An Arabidopsis Prenylated Rab Acceptor 1 Isoform, AtPRA1.B6, Displays Differential Inhibitory Effects on Anterograde Trafficking of Proteins at the Endoplasmic Reticulum

et al. 2011

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Prenylated Rab acceptors (PRAs), members of the Ypt-interacting protein family of small membrane proteins, are thought to aid the targeting of prenylated Rabs to their respective endomembrane compartments. In plants, the Arabidopsis (Arabidopsis thaliana) PRA1 family contains 19 members that display varying degrees of sequence homology to animal PRA1 and localize to the endoplasmic reticulum (ER) and/or endosomes. However, the exact role of these proteins remains to be fully characterized. In this study, the effect of AtPRA1.B6, a member of the AtPRA1 family, on the anterograde trafficking of proteins targeted to various endomembrane compartments was investigated. High levels of AtPRA1.B6 resulted in differential inhibition of coat protein complex II vesicle-mediated anterograde trafficking. The trafficking of the vacuolar proteins sporamin:GFP (for green fluorescent protein) and AALP:GFP, the secretory protein invertase:GFP, and the plasma membrane proteins PMP:GFP and H + -ATPase:GFP was inhibited in a dose-dependent manner, while the trafficking of the Golgi-localized proteins ST:GFP and KAM1(DC):mRFP was not affected. Conversely, in RNA interference plants displaying lower levels of AtPRA1.B6 transcripts, the trafficking efficiency of sporamin:GFP and AALP:GFP to the vacuole was increased. Localization and N-glycan pattern analyses of cargo proteins revealed that AtPRA1.B6-mediated inhibition of anterograde trafficking occurs at the ER. In addition, AtPRA1.B6 levels were controlled by cellular processes, including 26S proteasome-mediated proteolysis. Based on these results, we propose that AtPRA1. B6 is a negative regulator of coat protein complex II vesicle-mediated anterograde trafficking for a subset of proteins at the ER.

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“…Recent studies have demonstrated that protein export from the endoplasmic reticulum (ER) 2 is a selective process that is mediated through COPII-coated transport vesicles and dictated by short, linear sequences called ER export motifs (1)(2)(3)(4)(5)(6)(7)(8). COPII vesicles contain a small GTPase Sar1 and the heterodimeric Sec23/24 and Sec13/31 complexes.…”

mentioning

confidence: 99%

Di-acidic Motifs in the Membrane-distal C Termini Modulate the Transport of Angiotensin II Receptors from the Endoplasmic Reticulum to the Cell Surface

Zhang

Dong

et al. 2011

Journal of Biological Chemistry

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The molecular mechanisms underlying the endoplasmic reticulum (ER) export and cell surface transport of nascent G protein-coupled receptors (GPCRs) have just begun to be revealed and previous studies have shown that hydrophobic motifs in the putative amphipathic 8 th ␣-helical region within the membrane-proximal C termini play an important role. In this study, we demonstrate that di-acidic motifs in the membrane-distal, nonstructural C-terminal portions are required for the exit from the ER and transport to the plasma membrane of angiotensin II receptors, but not adrenergic receptors. More interestingly, distinct di-acidic motifs dictate optimal export trafficking of different angiotensin II receptors and export ability of each acidic residue in the di-acidic motifs cannot be fully substituted by other acidic residue. Moreover, the function of the di-acidic motifs is likely mediated through facilitating the recruitment of the receptors onto the ER-derived COPII transport vesicles. Therefore, the di-acidic motifs located in the membrane-distal C termini may represent the first linear motifs which recruit selective GPCRs onto the COPII vesicles to control their export from the ER.

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Role of Sec24 isoforms in selective export of membrane proteins from the endoplasmic reticulum

Cited by 168 publications

References 19 publications

The Organelle Proteome of the DT40 Lymphocyte Cell Line

The Organelle Proteome of the DT40 Lymphocyte Cell Line

An Arabidopsis Prenylated Rab Acceptor 1 Isoform, AtPRA1.B6, Displays Differential Inhibitory Effects on Anterograde Trafficking of Proteins at the Endoplasmic Reticulum

Di-acidic Motifs in the Membrane-distal C Termini Modulate the Transport of Angiotensin II Receptors from the Endoplasmic Reticulum to the Cell Surface

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