Brefeldin A-induced disassembly of the Golgi apparatus is followed by disruption of the endoplasmic reticulum in plant cells

1994 ◽  
Vol 45 (10) ◽  
pp. 1347-1351 ◽  
Author(s):  
Janey Henderson ◽  
Béatrice Stiat-Jeunemaitre ◽  
Richard Napier ◽  
Chris Hawes
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Plant Cells ◽  
Brefeldin A

scholarly journals cis-Golgi proteins accumulate near the ER exit sites and act as the scaffold for Golgi regeneration after brefeldin A treatment in tobacco BY-2 cells

2012 ◽  
Vol 23 (16) ◽  
pp. 3203-3214 ◽  
Author(s):  
Yoko Ito ◽  
Tomohiro Uemura ◽  
Keiko Shoda ◽  
Masaru Fujimoto ◽  
Takashi Ueda ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Fluorescent Proteins ◽  
Plant Cells ◽  
Brefeldin A ◽  
Eukaryotic Cells ◽  
Er Exit Sites ◽  
Intermediate Compartment ◽  
Golgi Protein ◽  
And Function

The Golgi apparatus forms stacks of cisternae in many eukaryotic cells. However, little is known about how such a stacked structure is formed and maintained. To address this question, plant cells provide a system suitable for live-imaging approaches because individual Golgi stacks are well separated in the cytoplasm. We established tobacco BY-2 cell lines expressing multiple Golgi markers tagged by different fluorescent proteins and observed their responses to brefeldin A (BFA) treatment and BFA removal. BFA treatment disrupted cis, medial, and trans cisternae but caused distinct relocalization patterns depending on the proteins examined. Medial- and trans-Golgi proteins, as well as one cis-Golgi protein, were absorbed into the endoplasmic reticulum (ER), but two other cis-Golgi proteins formed small punctate structures. After BFA removal, these puncta coalesced first, and then the Golgi stacks regenerated from them in the cis-to-trans order. We suggest that these structures have a property similar to the ER-Golgi intermediate compartment and function as the scaffold of Golgi regeneration.

scholarly journals Capacity of the Golgi Apparatus for Biogenesis from the Endoplasmic Reticulum

2003 ◽  
Vol 14 (12) ◽  
pp. 5011-5018 ◽  
Author(s):  
Sapna Puri ◽  
Adam D. Linstedt
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Self Assembly ◽  
De Novo ◽  
Brefeldin A ◽  
The Other ◽  
Matrix Proteins ◽  
Er Export ◽  
Golgi Matrix

It is unclear whether the mammalian Golgi apparatus can form de novo from the ER or whether it requires a preassembled Golgi matrix. As a test, we assayed Golgi reassembly after forced redistribution of Golgi matrix proteins into the ER. Two conditions were used. In one, ER redistribution was achieved using a combination of brefeldin A (BFA) to cause Golgi collapse and H89 to block ER export. Unlike brefeldin A alone, which leaves matrix proteins in relatively large remnant structures outside the ER, the addition of H89 to BFA-treated cells caused ER accumulation of all Golgi markers tested. In the other, clofibrate treatment induced ER redistribution of matrix and nonmatrix proteins. Significantly, Golgi reassembly after either treatment was robust, implying that the Golgi has the capacity to form de novo from the ER. Furthermore, matrix proteins reemerged from the ER with faster ER exit rates. This, together with the sensitivity of BFA remnants to ER export blockade, suggests that presence of matrix proteins in BFA remnants is due to cycling via the ER and preferential ER export rather than their stable assembly in a matrix outside the ER. In summary, the Golgi apparatus appears capable of efficient self-assembly.

scholarly journals FINE STRUCTURE AND ORGANELLE ASSOCIATIONS IN BROWN ALGAE

1965 ◽  
Vol 26 (2) ◽  
pp. 523-537 ◽  
Author(s):  
G. Benjamin Bouck
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Fine Structure ◽  
Golgi Apparatus ◽  
Nuclear Envelope ◽  
Brown Algae ◽  
Algal Cell ◽  
Plant Cells ◽  
Membrane Systems ◽  
Two Envelopes

The structural interrelationships among several membrane systems in the cells of brown algae have been examined by electron microscopy. In the brown algae the chloroplasts are surrounded by two envelopes, the outer of which in some cases is continuous with the nuclear envelope. The pyrenoid, when present, protrudes from the chloroplast, is also surrounded by the two chloroplast envelopes, and, in addition, is capped by a third dilated envelope or "pyrenoid sac." The regular apposition of the membranes around the pyrenoid contrasts with their looser appearance over the remainder of the chloroplast. The Golgi apparatus is closely associated with the nuclear envelope in all brown algae examined, but in the Fucales this association may extend to portions of the cytoplasmic endoplasmic reticulum as well. Evidence is presented for the derivation of vesicles, characteristic of those found in the formative region of the Golgi apparatus, from portions of the underlying nuclear envelope. The possibility that a structural channeling system for carbohydrate reserves and secretory precursors may be present in brown algae is considered. Other features of the brown algal cell, such as crystal-containing bodies, the variety of darkly staining vacuoles, centrioles, and mitochondria, are examined briefly, and compared with similar structures in other plant cells.

scholarly journals Transport of Bacterial Lipopolysaccharide to the Golgi Apparatus

1999 ◽  
Vol 190 (4) ◽  
pp. 523-534 ◽  
Author(s):  
Nathalie Thieblemont ◽  
Samuel D. Wright
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Hela Cells ◽  
Brefeldin A ◽  
Cellular Responses ◽  
Epithelial Cell Line ◽  
Cholera Toxin B Subunit ◽  
B Subunit ◽  
Cell Line Hela ◽  
Tetramethylrhodamine Isothiocyanate

Addition of lipopolysaccharide (LPS) to cells in the form of LPS–soluble (s)CD14 complexes induces strong cellular responses. During this process, LPS is delivered from sCD14 to the plasma membrane, and the cell-associated LPS is then rapidly transported to an intracellular site. This transport appears to be important for certain cellular responses to LPS, as drugs that block transport also inhibit signaling and cells from LPS-hyporesponsive C3H/HeJ mice fail to exhibit this transport. To identify the intracellular destination of fluorescently labeled LPS after its delivery from sCD14 into cells, we have made simultaneous observations of different organelles using fluorescent vital dyes or probes. Endosomes, lysosomes, the endoplasmic reticulum, and the Golgi apparatus were labeled using Texas red (TR)–dextran, LysoTracker™ Red DND-99, DiOC6(3), and boron dipyrromethane (BODIPY)–ceramide, respectively. After 30 min, LPS did not colocalize with endosomes, lysosomes, or endoplasmic reticulum in polymorphonuclear leukocytes, although some LPS-positive vesicles overlapped with the endosomal marker, fluorescent dextran. On the other hand, LPS did appear to colocalize with two markers of the Golgi apparatus, BODIPY–ceramide and TRITC (tetramethylrhodamine isothiocyanate)–labeled cholera toxin B subunit. We further confirmed the localization of LPS in the Golgi apparatus using an epithelial cell line, HeLa, which responds to LPS–sCD14 complexes in a CD14-dependent fashion: BODIPY–LPS was internalized and colocalized with fluorescently labeled Golgi apparatus probes in live HeLa cells. Morphological disruption of the Golgi apparatus in brefeldin A–treated HeLa cells caused intracellular redistribution of fluorescent LPS. These results are consistent with the Golgi apparatus being the primary delivery site of monomeric LPS.

scholarly journals Dynein Supports Motility of Endoplasmic Reticulum in the FungusUstilago maydis

2002 ◽  
Vol 13 (3) ◽  
pp. 965-977 ◽  
Author(s):  
Roland Wedlich-Söldner ◽  
Irene Schulz ◽  
Anne Straube ◽  
Gero Steinberg
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Intracellular Transport ◽  
Fluorescent Protein ◽  
Brefeldin A ◽  
Cytoplasmic Dynein ◽  
Organelle Transport ◽  
Minor Importance ◽  
Dependent Transport

The endoplasmic reticulum (ER) of most vertebrate cells is spread out by kinesin-dependent transport along microtubules, whereas studies in Saccharomyces cerevisiae indicated that motility of fungal ER is an actin-based process. However, microtubules are of minor importance for organelle transport in yeast, but they are crucial for intracellular transport within numerous other fungi. Herein, we set out to elucidate the role of the tubulin cytoskeleton in ER organization and dynamics in the fungal pathogen Ustilago maydis. An ER-resident green fluorescent protein (GFP)-fusion protein localized to a peripheral network and the nuclear envelope. Tubules and patches within the network exhibited rapid dynein-driven motion along microtubules, whereas conventional kinesin did not participate in ER motility. Cortical ER organization was independent of microtubules or F-actin, but reformation of the network after experimental disruption was mediated by microtubules and dynein. In addition, a polar gradient of motile ER-GFP stained dots was detected that accumulated around the apical Golgi apparatus. Both the gradient and the Golgi apparatus were sensitive to brefeldin A or benomyl treatment, suggesting that the gradient represents microtubule-dependent vesicle trafficking between ER and Golgi. Our results demonstrate a role of cytoplasmic dynein and microtubules in motility, but not peripheral localization of the ER inU. maydis.

scholarly journals Disruption of the Golgi Apparatus and Contribution of the Endoplasmic Reticulum to the SARS-CoV-2 Replication Complex

Viruses ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1798
Author(s):  
Ted Hackstadt ◽  
Abhilash I. Chiramel ◽  
Forrest H. Hoyt ◽  
Brandi N. Williamson ◽  
Cheryl A. Dooley ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Rna Synthesis ◽  
Light And Electron Microscopy ◽  
Brefeldin A ◽  
Viral Proteins ◽  
Protein Markers ◽  
Replication Complex ◽  
Golgi Fragmentation

A variety of immunolabeling procedures for both light and electron microscopy were used to examine the cellular origins of the host membranes supporting the SARS-CoV-2 replication complex. The endoplasmic reticulum has long been implicated as a source of membrane for the coronavirus replication organelle. Using dsRNA as a marker for sites of viral RNA synthesis, we provide additional evidence supporting ER as a prominent source of membrane. In addition, we observed a rapid fragmentation of the Golgi apparatus which is visible by 6 h and complete by 12 h post-infection. Golgi derived lipid appears to be incorporated into the replication organelle although protein markers are dispersed throughout the infected cell. The mechanism of Golgi disruption is undefined, but chemical disruption of the Golgi apparatus by brefeldin A is inhibitory to viral replication. A search for an individual SARS-CoV-2 protein responsible for this activity identified at least five viral proteins, M, S, E, Orf6, and nsp3, that induced Golgi fragmentation when expressed in eukaryotic cells. Each of these proteins, as well as nsp4, also caused visible changes to ER structure as shown by correlative light and electron microscopy (CLEM). Collectively, these results imply that specific disruption of the Golgi apparatus is a critical component of coronavirus replication.

scholarly journals Cytosolic Free N-Glycans Are Retro-Transported Into the Endoplasmic Reticulum in Plant Cells

2021 ◽  
Vol 11 ◽  
Author(s):  
Makoto Katsube ◽  
Natsuki Ebara ◽  
Megumi Maeda ◽  
Yoshinobu Kimura
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Folding ◽  
Golgi Apparatus ◽  
Plant Cells ◽  
Complex Type ◽  
Secretion Pathway ◽  
Amyloid Fibril Formation ◽  
Free Oligosaccharides ◽  
Β Amyloid ◽  
Protein Secretion Pathway

During endoplasmic reticulum (ER)-associated degradation, free N-glycans (FNGs) are produced from misfolded nascent glycoproteins via the combination of the cytosolic peptide N-glycanase (cPNGase) and endo-β-N-acetylglucosaminidase (ENGase) in the plant cytosol. The resulting high-mannose type (HMT)-FNGs, which carry one GlcNAc residue at the reducing end (GN1-FNGs), are ubiquitously found in developing plant cells. In a previous study, we found that HMT-FNGs assisted in protein folding and inhibited β-amyloid fibril formation, suggesting a possible biofunction of FNGs involved in the protein folding system. However, whether these HMT-FNGs occur in the ER, an organelle involved in protein folding, remained unclear. On the contrary, we also reported the presence of plant complex type (PCT)-GN1-FNGs, which carry the Lewisa epitope at the non-reducing end, indicating that these FNGs had been fully processed in the Golgi apparatus. Since plant ENGase was active toward HMT-N-glycans but not PCT-N-glycans that carry β1-2xylosyl and/or α1-3 fucosyl residue(s), these PCT-GN1-FNGs did not appear to be produced from fully processed glycoproteins that harbored PCT-N-glycans via ENGase activity. Interestingly, PCT-GN1-FNGs were found in the extracellular space, suggesting that HMT-GN1-FNGs formed in the cytosol might be transported back to the ER and processed in the Golgi apparatus through the protein secretion pathway. As the first step in elucidating the production mechanism of PCT-GN1-FNGs, we analyzed the structures of free oligosaccharides in plant microsomes and proved that HMT-FNGs (Man9-7GlcNAc1 and Man9-8GlcNAc2) could be found in microsomes, which almost consist of the ER compartments.

Brefeldin A affects the endomembrane system and vesicle trafficking in higher plants

1993 ◽  
Vol 51 ◽  
pp. 192-193
Author(s):  
Béatrice Satiat-Jeunemaitre ◽  
Jancy Henderson ◽  
David Evans ◽  
Kim Crooks ◽  
Mark Fricker ◽  
...  
Keyword(s):  
Golgi Apparatus ◽  
Higher Plants ◽  
Fatty Acid Derivative ◽  
Plant Cells ◽  
Brefeldin A ◽  
Endomembrane System ◽  
Animal Cells ◽  
Higher Plant ◽  
Membrane Flow ◽  
Golgi Stack

In plant cells, as in animal cells, many macromolecules and membranes are transported by vesicle vectors through both the exocytotic and endocytotic pathways. In order to elucidate the mechanisms and molecular events of such trafficking we are using a set of drugs known to perturb membrane flow in plant cells in combination with immunocytochemical studies using a bank of monoclonal antibodies to various components of the endomembrane system and cell surface. In animal cells, one such drug, Brefeldin A, a fungal fatty acid derivative which causes disruption of the Golgi apparatus, has recently been used as a tool to dissect the mechanisms of vesicle flow from the endoplasmic reticulum to the Golgi apparatus and down the cisternae of the Golgi stack (1). It has been demonstrated that BFA also has a dramatic effect on the Golgi apparatus in higher plant cells (2,3,4).In this paper we report on recent work on the disruption of the plant Golgi apparatus with BFA and the redistribution of endomembrane marker epitopes after drug treatment of roots and suspension culture cells.

Complex envelope system. Membrane systems of plant cells

1974 ◽  
Vol 268 (891) ◽  
pp. 119-128 ◽  
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Smooth Endoplasmic Reticulum ◽  
Plant Cells ◽  
Membrane System ◽  
Cellulose Microfibrils ◽  
Membrane Systems ◽  
Complex Envelope ◽  
Pectic Substances ◽  
The Matrix

The membrane system is made up of the nuclear envelopes, rough and smooth endoplasmic reticulum, Golgi apparatus and plasmalemma. Interconnexions between the various parts of the system are shown and these probably represent a flow of membrane from the endoplasmic reticulum through the Golgi apparatus to the plasmalemma. Membrane fractions have been isolated from broken cells and their function in the synthesis of polysaccharides established. It has been shown that the matrix polysaccharides of the wall (pectic substances and hemicelluloses) are formed within the membranes and that the pattern of synthesis of these polymers changes during differentiation of the cells. Cellulose microfibrils are probably synthesized at the plasmalemma which is formed by incorporation of membrane bounded vesicles from the Golgi apparatus. Thus the assembly of the polymers takes place either when the membrane is within the cytoplasm or when it is incorporated as the plasmalemma of the cell.

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