Intercompartmental transport in the Golgi complex is a dissociative process: facile transfer of membrane protein between two Golgi populations.

Rothman, James E.; Miller, R L; Urbani, Lenore J.

doi:10.1083/jcb.99.1.260

Cited by 146 publications

(51 citation statements)

References 27 publications

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“…The vesicles which mediate the traffic of the secretory pathway can diffuse throughout the cytosol prior to being consumed at their target organelle (Novick and Schekman, 1979;Rothman et al, 1984) and this traffic continues throughout interphase as long as the donor and target organelle are intact. Vacuole inheritance is also mediated by tubulovesicular traffic (Weisman and Wickner, 1988;Raymond et al, 1992;Gomes de Mesquita et al, 1991) but the bud vacuole is founded at a precise time in the cell cycle by a spatially constrained stream of vesicles and membranous tubules (Weisman and Wickner, 1988).…”

Section: Discussionmentioning

confidence: 99%

Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae.

Wickner

1996

The Journal of Cell Biology

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The vacuole of Saccharomyces cerevisiae projects a stream of tubules a and vesicles (a "segregation structure") into the bud in early S phase. We have described an in vitro reaction, requiring physiological temperature, ATP, and cytosol, in which isolated vacuoles form segregation structures and fuse. This in vitro reaction is defective when reaction components are prepared from vac mutants that are defective in this process in vivo, Fractionation of the cytosol reveals at least three components, each of which can support the vacuole fusion reaction, and two stimulatory fractions. Purification of one "low molecular weight activity" (LMA1) yields a heterodimeric protein with a thioredoxin subunit. Most of the thioredoxin of yeast is in this complex rather than the well-studied monomer. A deletion of both S. cerevisiae thioredoxin genes causes a striking vacuole inheritance defect in vivo, establishing a role for thioredoxin as a novel factor in this trafficking reaction.

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Section: Discussionmentioning

confidence: 99%

Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae.

Wickner

1996

The Journal of Cell Biology

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“…However, it is also possible that the fragmentation of the Golgi apparatus that occurs in mitosis represents unproductive fusion events. Rothman et al (34) have shown that productive transfer from the Golgi apparatus of one cell to that of another can occur after fusion, so the simple physical separation of Golgi elements that occurs during mitosis need not block transfer. However, the organization of protein and other elements within a Golgi stack or saccule presumably was preserved in the cell fusion experiments, whereas the fragmentation during mitosis could create vesicles that are structurally disorganized or restricted in enzymatic function, thus further reducing the efficiency of productive transfer, or perhaps affecting recognition signals that optimize productive fusions.…”

Section: Discussionmentioning

confidence: 99%

Glycosaminoglycan synthesis is depressed during mitosis and elevated during early G1.

Preston¹,

Regula²,

Sager³

et al. 1985

The Journal of Cell Biology

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[35S]Sulfate incorporation was measured in populations of Chinese hamster ovary cells enriched for mitotics, early G1 cells, and interphase monolayers or suspensions. Incorporation was determined by biochemical analysis of extracts and quantitative autoradiography of thick sections. 90% of [35S]sulfate was incorporated into glycosaminoglycan (GAG). Incorporation was depressed fourfold in mitotics and stimulated by from two-to three-fold in early G1 cells relative to mixed interphase cells. GAG synthesis was maintained into late G2. Thus, the rate of GAG biosynthesis was correlated temporally with the detachment and reattachment of cells to substrate.Inhibitors of protein synthesis brought about the rapid arrest of GAG biosynthesis. However, xylosides, which bypass the requirement for core protein, did not bring oligosaccharide sulfation in mitotics to interphase levels. These observations indicate an inhibition of Golgi processing and are consistent with a generalized defect of membrane vesicle-mediated transport during mitosis.In their pioneering work on glycosaminoglycan (GAG) ~ synthesis by cultured cells, Kraemer and Tobey (14) showed that a significant fraction of trypsin-releasable (i.e., surface) GAGs was shed during mitosis. Although the Chinese hamster ovary (CHO) cells analyzed were grown in suspension, viewed from the perspective of recent studies of cell adhesion (e.g., reference 32), it seemed likely that the shedding of GAGs was related to the familiar ease with which mitotic cells are detached from monolayers. We hypothesized that any effect of shedding would be reinforced by a coordinate reduction in GAG synthesis that would delay replacement of surface GAGs, and that a rapid resumption of GAG synthesis in early G i would be required for cell reattachment.A parallel motivation for our studies arose from the accumulating evidence that membrane vesicle-mediated transport is depressed during mitosis. This is shown clearly in work from our laboratory demonstrating depression of endocytosis ( 1, 2) and transferrin recycling (35) and from that of Warren and colleagues showing a decrease in surface transferrin receptors (41), and in depression of histamine secretion (9). These studies focus primarily on fusion and budding events at the plasma membrane. In addition, the insertion of the G t Abbreviations used in this paper. CHO, Chinese hamster ovary; GAG, glycosaminoglycan.protein of vesicular stomatitis virus is also depressed during mitosis (42), which suggests a defect in its processing through the Golgi apparatus. Since carbohydrate chains of GAGs are largely assembled within the Golgi apparatus by multiple enzymes (31), a general defect in processing should affect GAG synthesis. Moreover, the apparent localization of sulfation to the trans-Gol~ apparatus (7, 44) would serve to focus on later steps of the processing sequence.We describe here the rates of labeled sulfate incorporation into CHO cells during mitosis, in their subsequent movement into G1, and in late G2. We followed incorporat...

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“…Megalocytosis began to develop within 12-18 h and was fully evident by 24 -48 h. Most experiments were performed 4 days after addition of MCTP. In cell fractionation experiments, to avoid detecting proteins that were merely transiting through the Golgi at the time of harvesting the cells, and to thus isolate only the Golgi resident proteins, cultures were pretreated with cycloheximide (10 g/ml) for 2 h (48,53). Cycloheximide inhibits protein synthesis and subsequent entry of any new proteins into the ER-Golgi and thus flushes away secretable proteins from the Golgi (48,53).…”

Section: Methodsmentioning

confidence: 99%

Dysfunction of Golgi tethers, SNAREs, and SNAPs in monocrotaline-induced pulmonary hypertension

Sehgal¹,

Mukhopadhyay²,

Xu³

et al. 2007

American Journal of Physiology-Lung Cellular and Molecular Phys

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Monocrotaline (MCT)-induced pulmonary hypertension (PH) in the rat is a widely used experimental model. We have previously shown that MCT pyrrole (MCTP) produces loss of caveolin-1 (cav-1) and endothelial nitric oxide synthase from plasma membrane raft microdomains in pulmonary arterial endothelial cells (PAEC) with the trapping of these proteins in the Golgi organelle (the Golgi blockade hypothesis). In the present study, we investigated the mechanisms underlying this intracellular trafficking block in experiments in cell culture and in the MCT-treated rat. In cell culture, PAEC showed trapping of cav-1 in Golgi membranes as early as 6 h after exposure to MCTP. Phenotypic megalocytosis and a reduction in anterograde trafficking (assayed in terms of the secretion of horseradish peroxidase derived from exogenously transfected expression constructs) were evident within 12 h after MCTP. Cell fractionation and immunofluorescence techniques revealed the marked accumulation of diverse Golgi tethers, soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptors (SNAREs), and soluble NSF attachment proteins (SNAPs), which mediate membrane fusion during vesicular trafficking (GM130, p115, giantin, golgin 84, clathrin heavy chain, syntaxin-4, -6, Vti1a, Vti1b, GS15, GS27, GS28, SNAP23, and ␣-SNAP) in the enlarged/ circumnuclear Golgi in MCTP-treated PAEC and A549 lung epithelial cells. Moreover, NSF, an ATPase required for the "disassembly" of SNARE complexes subsequent to membrane fusion, was increasingly sequestered in non-Golgi membranes. Immunofluorescence studies of lung tissue from MCT-treated rats confirmed enlargement of perinuclear Golgi elements in lung arterial endothelial and parenchymal cells as early as 4 days after MCT. Thus MCTinduced PH represents a disease state characterized by dysfunction of Golgi tethers, SNAREs, and SNAPs and of intracellular vesicular trafficking.soluble N-ethylmaleimide-sensitive factor attachment protein receptors; endothelium; intracellular membrane trafficking; Golgi blockade PULMONARY HYPERTENSION (PH) is a progressive disease with high morbidity and mortality (10). As the clinical manifestations of PH typically occur long after the initial injury, experimental models provide an opportunity to investigate the initiating mechanism at the cellular and biochemical levels (45). Among experimental models of PH, administration of the pyrrolizidine alkaloid monocrotaline (MCT) to the juvenile male rat has and continues to be used extensively (21,29). In this model, progressive PH develops 10 -14 days after a single injection of MCT. The injected MCT is converted to its active pyrrolic derivative [MCT pyrrole (MCTP)] by the cytochrome P-450 system in the liver (36). The bioactive MCTP, which has a short half-life (ϳ3 s in aqueous medium), affects the first vascular bed it encounters: the pulmonary arterial system. The major cellular targets of MCT in the rat lung include the pulmonary arterial endothelial cells (PAEC), pulmonary arterial smooth muscle cells (PASMC), and ...

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Intercompartmental transport in the Golgi complex is a dissociative process: facile transfer of membrane protein between two Golgi populations.

Abstract: The transfer of the vesicular stomatitis virus-encoded glycoprotein (G protein) between Golgi populations in fused cells (Rothman, J . E., L. J . Urbani, and R . Brands . 1984 .

Cited by 146 publications

References 27 publications

Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae.

Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae.

Glycosaminoglycan synthesis is depressed during mitosis and elevated during early G1.

Dysfunction of Golgi tethers, SNAREs, and SNAPs in monocrotaline-induced pulmonary hypertension

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