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Peri-Golgi vesicles contain retrograde but not anterograde proteins consistent with the cisternal progression model of intra-Golgi transport.

Martinez-Menárguez JA, Prekeris R, Oorschot VM, Scheller R, Slot JW, Geuze HJ, Klumperman J - J. Cell Biol. (2001)

Bottom Line: A cisternal progression mode of intra-Golgi transport requires that Golgi resident proteins recycle by peri-Golgi vesicles, whereas the alternative model of vesicular transport predicts anterograde cargo proteins to be present in such vesicles.We found significant levels of the Golgi resident enzyme mannosidase II and the transport machinery proteins giantin, KDEL-receptor, and rBet1 in coatomer protein I-coated cisternal rims and peri-Golgi vesicles.By contrast, when cells expressed vesicular stomatitis virus protein G this anterograde marker was largely absent from the peri-Golgi vesicles.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, School of Medicine, University of Murcia, 30071 Murcia, Spain.

ABSTRACT
A cisternal progression mode of intra-Golgi transport requires that Golgi resident proteins recycle by peri-Golgi vesicles, whereas the alternative model of vesicular transport predicts anterograde cargo proteins to be present in such vesicles. We have used quantitative immuno-EM on NRK cells to distinguish peri-Golgi vesicles from other vesicles in the Golgi region. We found significant levels of the Golgi resident enzyme mannosidase II and the transport machinery proteins giantin, KDEL-receptor, and rBet1 in coatomer protein I-coated cisternal rims and peri-Golgi vesicles. By contrast, when cells expressed vesicular stomatitis virus protein G this anterograde marker was largely absent from the peri-Golgi vesicles. These data suggest a role of peri-Golgi vesicles in recycling of Golgi residents, rather than an important role in anterograde transport.

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KDELr (10 nm gold) and rBet1 (15 nm gold) colocalize in peripheral VTCs (pVTC) but not peri-Golgi vesicles. (A) High levels of KDELr and rBet1 are found in membranes of a peripheral VTC. (B) In the Golgi (G) region, rBet1 is present in irregularly shaped VTC membranes but absent from peri-Golgi vesicles (arrows). P, plasma membrane. Bars, 200 nm.
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fig7: KDELr (10 nm gold) and rBet1 (15 nm gold) colocalize in peripheral VTCs (pVTC) but not peri-Golgi vesicles. (A) High levels of KDELr and rBet1 are found in membranes of a peripheral VTC. (B) In the Golgi (G) region, rBet1 is present in irregularly shaped VTC membranes but absent from peri-Golgi vesicles (arrows). P, plasma membrane. Bars, 200 nm.

Mentions: Finally, we analyzed the lateral distribution of the SNARE protein rBet1, which has been implicated in membrane fusion events early after the formation of ER transport vesicles (Hay et al., 1997, 1998; Zhang et al., 1997; Chao et al., 1999). Consistent with previous studies, the bulk of rBet1 labeling was found in VTCs (Figs. 5 and 7). When double labeled with giantin, rBet1 clearly labeled a distinct population of membranes (Fig. 5). In the Golgi stack, rBet1 was present predominantly in G1 and G2 (Table I). Assessment of its lateral distribution within the Golgi complex showed that almost 60% of rBet1 located to COPI-coated buds and peri-Golgi vesicles (Table II).


Peri-Golgi vesicles contain retrograde but not anterograde proteins consistent with the cisternal progression model of intra-Golgi transport.

Martinez-Menárguez JA, Prekeris R, Oorschot VM, Scheller R, Slot JW, Geuze HJ, Klumperman J - J. Cell Biol. (2001)

KDELr (10 nm gold) and rBet1 (15 nm gold) colocalize in peripheral VTCs (pVTC) but not peri-Golgi vesicles. (A) High levels of KDELr and rBet1 are found in membranes of a peripheral VTC. (B) In the Golgi (G) region, rBet1 is present in irregularly shaped VTC membranes but absent from peri-Golgi vesicles (arrows). P, plasma membrane. Bars, 200 nm.
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Related In: Results  -  Collection

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fig7: KDELr (10 nm gold) and rBet1 (15 nm gold) colocalize in peripheral VTCs (pVTC) but not peri-Golgi vesicles. (A) High levels of KDELr and rBet1 are found in membranes of a peripheral VTC. (B) In the Golgi (G) region, rBet1 is present in irregularly shaped VTC membranes but absent from peri-Golgi vesicles (arrows). P, plasma membrane. Bars, 200 nm.
Mentions: Finally, we analyzed the lateral distribution of the SNARE protein rBet1, which has been implicated in membrane fusion events early after the formation of ER transport vesicles (Hay et al., 1997, 1998; Zhang et al., 1997; Chao et al., 1999). Consistent with previous studies, the bulk of rBet1 labeling was found in VTCs (Figs. 5 and 7). When double labeled with giantin, rBet1 clearly labeled a distinct population of membranes (Fig. 5). In the Golgi stack, rBet1 was present predominantly in G1 and G2 (Table I). Assessment of its lateral distribution within the Golgi complex showed that almost 60% of rBet1 located to COPI-coated buds and peri-Golgi vesicles (Table II).

Bottom Line: A cisternal progression mode of intra-Golgi transport requires that Golgi resident proteins recycle by peri-Golgi vesicles, whereas the alternative model of vesicular transport predicts anterograde cargo proteins to be present in such vesicles.We found significant levels of the Golgi resident enzyme mannosidase II and the transport machinery proteins giantin, KDEL-receptor, and rBet1 in coatomer protein I-coated cisternal rims and peri-Golgi vesicles.By contrast, when cells expressed vesicular stomatitis virus protein G this anterograde marker was largely absent from the peri-Golgi vesicles.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, School of Medicine, University of Murcia, 30071 Murcia, Spain.

ABSTRACT
A cisternal progression mode of intra-Golgi transport requires that Golgi resident proteins recycle by peri-Golgi vesicles, whereas the alternative model of vesicular transport predicts anterograde cargo proteins to be present in such vesicles. We have used quantitative immuno-EM on NRK cells to distinguish peri-Golgi vesicles from other vesicles in the Golgi region. We found significant levels of the Golgi resident enzyme mannosidase II and the transport machinery proteins giantin, KDEL-receptor, and rBet1 in coatomer protein I-coated cisternal rims and peri-Golgi vesicles. By contrast, when cells expressed vesicular stomatitis virus protein G this anterograde marker was largely absent from the peri-Golgi vesicles. These data suggest a role of peri-Golgi vesicles in recycling of Golgi residents, rather than an important role in anterograde transport.

Show MeSH
Related in: MedlinePlus