An Arabidopsis thaliana cDNA Complements the N-Acetylglucosaminyltransferase I Deficiency of CHO Lec1 Cells

Bakker, Hans; Lommen, Arjen; Jordi, W.; Stiekema, Willem J.; Bosch, Dirk

doi:10.1006/bbrc.1999.1117

Cited by 28 publications

(20 citation statements)

References 22 publications

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“…Indeed, the first glycosyltransferases specific for plant N-glycan maturation, N-acetylglucosaminyltransferase I (GnTI) and ␣-(1,3)-fucosyltransferase, were recently cloned from tobacco, Arabidopsis, and mung bean (Bakker et al, 1999;Leiter et al, 1999;Strasser et al, 1999), and we have provided evidence for Golgi localization of GnTI (Essl et al, 1999). However, detailed immunolocalization by electron microscopy is not available for any glycosyltransferase responsible for plant N-glycan maturation.…”

Section: The "All In a Single Bag" Theory For Plant N-glycan Maturationmentioning

confidence: 92%

Protein Recycling from the Golgi Apparatus to the Endoplasmic Reticulum in Plants and Its Minor Contribution to Calreticulin Retention

et al. 2000

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Using pulse-chase experiments combined with immunoprecipitation and N-glycan structural analysis, we showed that the retrieval mechanism of proteins from post-endoplasmic reticulum (post-ER) compartments is active in plant cells at levels similar to those described previously for animal cells. For instance, recycling from the Golgi apparatus back to the ER is sufficient to block the secretion of as much as 90% of an extracellular protein such as the cell wall invertase fused with an HDEL C-terminal tetrapeptide. Likewise, recycling can sustain fast retrograde transport of Golgi enzymes into the ER in the presence of brefeldin A. However, on the basis of our data, we propose that this retrieval mechanism in plants has little impact on the ER retention of a soluble ER protein such as calreticulin. Indeed, the latter is retained in the ER without any N-glycan-related evidence for a recycling through the Golgi apparatus. Taken together, these results indicate that calreticulin and perhaps other plant reticuloplasmins are possibly largely excluded from vesicles exported from the ER. Instead, they are probably retained in the ER by mechanisms that rely primarily on signals other than H/KDEL motifs. INTRODUCTIONIn eukaryotic cells, most proteins that reside in the endoplasmic reticulum (ER) contain information in their primary structure that determines their subcellular localization. Keeping these proteins in the ER can be achieved either by strict retention in this particular compartment or by retrieval and retrograde transport back to the ER when they leave the ER (reviewed in Gomord et al., 1999;Pagny et al., 1999; Vitale and Denecke, 1999). For soluble ER resident proteins or soluble reticuloplasmins, ER residency depends largely on the presence of the specific tetrapeptide H/KDEL-or closely related sequences-at their C termini (Munro and Pelham, 1987;Andres et al., 1990). Use of different tetrapeptide sequences fused to non-ER secretory proteins as reporter probes has shown that these signals are sufficient for retention of soluble proteins in the ER (Rose and Doms, 1988;Herman et al., 1990; Denecke et al., 1992; Wandelt et al., 1992; Boevink et al., 1996;Gomord et al., 1997). Moreover, for yeast and mammalian cells, the H/KDEL-dependent retention mechanism has been proposed to promote a continual recycling of ER resident proteins whenever they exit the ER and are secreted into a post-ER compartment (Pelham, 1988).Arguing for such a recycling of ER proteins is the observation that reporter glycoproteins fused with H/KDEL motifs undergo specific N-glycan modifications. Indeed, the latter are markers of post-ER events. For example, in animal cells, the lysosomal glycoprotein cathepsin D, when modified by the addition of a KDEL C-terminal extension, accumulates in the ER even though it carries N-glycans containing the N -acetylglucosamine 1-phosphate residues that are typical of passage through the cis Golgi compartment (Pelham, 1988). Some post-ER glycan maturations have also been found on reporter glycoproteins ...

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Section: The "All In a Single Bag" Theory For Plant N-glycan Maturationmentioning

confidence: 92%

Protein Recycling from the Golgi Apparatus to the Endoplasmic Reticulum in Plants and Its Minor Contribution to Calreticulin Retention

et al. 2000

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“…A cytoplasmic form of human UXS1 (cytUXS1) was amplified from the full-length clone by primers amplifying the region coding from amino acid 38 to the C terminus and cloned in pcDNA3-FLAGhemagglutinin like the full-length construct. Arabidopsis thaliana UXS3 (30) (AT5G59290) was amplified by primers covering the complete open reading frame from a cDNA library (31) and cloned into pcDNA3-FLAG-hemagglutinin. Arabidopsis ␤1,2-xylosyltransferase (AtXylT; AT5G55500) (32) was isolated from the same cDNA library using a sibling selection procedure with PCR primers based on a partial soybean XylT sequence (33).…”

Section: Methodsmentioning

confidence: 99%

Functional UDP-xylose Transport across the Endoplasmic Reticulum/Golgi Membrane in a Chinese Hamster Ovary Cell Mutant Defective in UDP-xylose Synthase

Bakker

Oka

Ashikov

et al. 2009

Journal of Biological Chemistry

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In mammals, xylose is found as the first sugar residue of the tetrasaccharide GlcA␤1-3Gal␤1-3Gal␤1-4Xyl␤1-O-Ser, initiating the formation of the glycosaminoglycans heparin/heparan sulfate and chondroitin/dermatan sulfate. It is also found in the trisaccharide Xyl␣1-3Xyl␣1-3Glc␤1-O-Ser on epidermal growth factor repeats of proteins, such as Notch. UDP-xylose synthase (UXS), which catalyzes the formation of the UDP-xylose substrate for the different xylosyltransferases through decarboxylation of UDP-glucuronic acid, resides in the endoplasmic reticulum and/or Golgi lumen. Since xylosylation takes place in these organelles, no obvious requirement exists for membrane transport of UDP-xylose. However, UDP-xylose transport across isolated Golgi membranes has been documented, and we recently succeeded with the cloning of a human UDP-xylose transporter (SLC25B4). Here we provide new evidence for a functional role of UDP-xylose transport by characterization of a new Chinese hamster ovary cell mutant, designated pgsI-208, that lacks UXS activity. The mutant fails to initiate glycosaminoglycan synthesis and is not capable of xylosylating Notch. Complementation was achieved by expression of a cytoplasmic variant of UXS, which proves the existence of a functional Golgi UDP-xylose transporter. A ϳ200 fold increase of UDP-glucuronic acid occurred in pgsI-208 cells, demonstrating a lack of UDP-xylose-mediated control of the cytoplasmically localized UDP-glucose dehydrogenase in the mutant. The data presented in this study suggest the bidirectional transport of UDP-xylose across endoplasmic reticulum/Golgi membranes and its role in controlling homeostasis of UDP-glucuronic acid and UDP-xylose production.

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“…Twenty-four predicted glycosyltransferases cited in CAZy (27) are localized to the Golgi apparatus, reflecting the fact that this organelle is specialized for glycosylation. These glycosyl transferases include enzymes involved in N-glycosylation, such as Nacetyl glucosaminyl transferase I, and cell wall biosynthetic enzymes, such as Quasimodo1 (28,29). Interestingly, the cellulose synthase proteins CesA1 and CesA3 also are localized to the Golgi apparatus (30).…”

Section: Ermentioning

confidence: 99%

Mapping the Arabidopsis organelle proteome

Dunkley

Hester

Shadforth

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

507

428

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A challenging task in the study of the secretory pathway is the identification and localization of new proteins to increase our understanding of the functions of different organelles. Previous proteomic studies of the endomembrane system have been hindered by contaminating proteins, making it impossible to assign proteins to organelles. Here we have used the localization of organelle proteins by the isotope tagging technique in conjunction with isotope tags for relative and absolute quantitation and 2D liquid chromatography for the simultaneous assignment of proteins to multiple subcellular compartments. With this approach, the density gradient distributions of 689 proteins from Arabidopsis thaliana were determined, enabling confident and simultaneous localization of 527 proteins to the endoplasmic reticulum, Golgi apparatus, vacuolar membrane, plasma membrane, or mitochondria and plastids. This parallel analysis of endomembrane components has enabled protein steady-state distributions to be determined. Consequently, genuine organelle residents have been distinguished from contaminating proteins and proteins in transit through the secretory pathway.endomembrane ͉ localization of organelle proteins by isotope tagging ͉ isotope tags for relative and absolute quantitation ͉ organelle proteomics P roteins are spatially organized according to their functions within the eukaryotic cell. Therefore, protein localization is an important step toward assigning functions to the thousands of uncharacterized proteins predicted by the genome-sequencing projects. Proteomics provides powerful tools for characterizing the protein contents of organelles. Confident protein localization, however, requires that either organelle preparations are free of contaminants or that techniques are used to discriminate between genuine organelle residents and contaminating proteins (1). Although reasonably pure preparations of some organelles, such as mitochondria, can be achieved, the components of the endomembrane system so far have proved recalcitrant to purification (2, 3). The constituent organelles of the endomembrane system have similar sizes and densities, making them difficult to separate. In addition, the proteins that reside within this system are in a constant state of flux. Endomembrane proteins traffic through the system en route to their final destination; for example, plasma membrane (PM) proteins travel although the endoplasmic reticulum (ER) and the Golgi apparatus before reaching the cell surface. Proteins within the endomembrane system also cycle between compartments; for example, ER residents continuously escape to the Golgi apparatus and are retrieved in COPI vesicles (4). Consequently, it is not sufficient merely to identify the proteins within a single organelle-enriched fraction. Instead, the steady-state distributions of proteins within the whole endomembrane system must be determined if a realistic insight into the subcellular localization of endomembrane proteins is to be achieved.Localization of organelle proteins by...

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An Arabidopsis thaliana cDNA Complements the N-Acetylglucosaminyltransferase I Deficiency of CHO Lec1 Cells

Cited by 28 publications

References 22 publications

Protein Recycling from the Golgi Apparatus to the Endoplasmic Reticulum in Plants and Its Minor Contribution to Calreticulin Retention

Protein Recycling from the Golgi Apparatus to the Endoplasmic Reticulum in Plants and Its Minor Contribution to Calreticulin Retention

Functional UDP-xylose Transport across the Endoplasmic Reticulum/Golgi Membrane in a Chinese Hamster Ovary Cell Mutant Defective in UDP-xylose Synthase

Mapping the Arabidopsis organelle proteome

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