Limits...
A novel role for dp115 in the organization of tER sites in Drosophila.

Kondylis V, Rabouille C - J. Cell Biol. (2003)

Bottom Line: However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays.The effects were specific for dp115.Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p.

View Article: PubMed Central - PubMed

Affiliation: The Wellcome Trust Center for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, UK.

ABSTRACT
Here, we describe that depletion of the Drosophila homologue of p115 (dp115) by RNA interference in Drosophila S2 cells led to important morphological changes in the Golgi stack morphology and the transitional ER (tER) organization. Using conventional and immunoelectron microscopy and confocal immunofluorescence microscopy, we show that Golgi stacks were converted into clusters of vesicles and tubules, and that the tERs (marked by Sec23p) lost their focused organization and were now dispersed throughout the cytoplasm. However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays. The effects were specific for dp115. Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p. Taken together, these studies suggest that dp115 could be implicated in the architecture of both the Golgi stacks and the tER sites.

Show MeSH

Related in: MedlinePlus

Effect of dp115 depletion on the organization of the tER sites. S2 cells were processed for confocal immunofluorescence microscopy using the anti-Sec23p antibody (green; A and D) and the d120kd antibody (red; B and E) in mock-depleted (+ds EGFP; A–C) and dp115-depleted cells (+ds dp115; D–F). Projections of 30 sections are presented, and in merge images (C and F), the overlap is yellow. Note that in mock-depleted cells, almost every dSec23p-positive structure is found in close proximity to a d120kd-positive one. Bar, 5 μm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2172793&req=5

fig6: Effect of dp115 depletion on the organization of the tER sites. S2 cells were processed for confocal immunofluorescence microscopy using the anti-Sec23p antibody (green; A and D) and the d120kd antibody (red; B and E) in mock-depleted (+ds EGFP; A–C) and dp115-depleted cells (+ds dp115; D–F). Projections of 30 sections are presented, and in merge images (C and F), the overlap is yellow. Note that in mock-depleted cells, almost every dSec23p-positive structure is found in close proximity to a d120kd-positive one. Bar, 5 μm.

Mentions: In immunofluorescence, these two antibodies gave similar patterns. The dSec23p pattern corresponded to 20 ± 8 large fluorescent objects dispersed in the cytoplasm (Fig. 6 A). The d120kd pattern corresponded to 18 ± 7 similar large fluorescent objects (Fig. 6 B). These two patterns overlapped partially (Fig. 6 C). By IEM, the region of overlap corresponded to the interface between the Golgi stack and the tER where the two antigens are in close proximity (Fig. 5 C).


A novel role for dp115 in the organization of tER sites in Drosophila.

Kondylis V, Rabouille C - J. Cell Biol. (2003)

Effect of dp115 depletion on the organization of the tER sites. S2 cells were processed for confocal immunofluorescence microscopy using the anti-Sec23p antibody (green; A and D) and the d120kd antibody (red; B and E) in mock-depleted (+ds EGFP; A–C) and dp115-depleted cells (+ds dp115; D–F). Projections of 30 sections are presented, and in merge images (C and F), the overlap is yellow. Note that in mock-depleted cells, almost every dSec23p-positive structure is found in close proximity to a d120kd-positive one. Bar, 5 μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2172793&req=5

fig6: Effect of dp115 depletion on the organization of the tER sites. S2 cells were processed for confocal immunofluorescence microscopy using the anti-Sec23p antibody (green; A and D) and the d120kd antibody (red; B and E) in mock-depleted (+ds EGFP; A–C) and dp115-depleted cells (+ds dp115; D–F). Projections of 30 sections are presented, and in merge images (C and F), the overlap is yellow. Note that in mock-depleted cells, almost every dSec23p-positive structure is found in close proximity to a d120kd-positive one. Bar, 5 μm.
Mentions: In immunofluorescence, these two antibodies gave similar patterns. The dSec23p pattern corresponded to 20 ± 8 large fluorescent objects dispersed in the cytoplasm (Fig. 6 A). The d120kd pattern corresponded to 18 ± 7 similar large fluorescent objects (Fig. 6 B). These two patterns overlapped partially (Fig. 6 C). By IEM, the region of overlap corresponded to the interface between the Golgi stack and the tER where the two antigens are in close proximity (Fig. 5 C).

Bottom Line: However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays.The effects were specific for dp115.Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p.

View Article: PubMed Central - PubMed

Affiliation: The Wellcome Trust Center for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, UK.

ABSTRACT
Here, we describe that depletion of the Drosophila homologue of p115 (dp115) by RNA interference in Drosophila S2 cells led to important morphological changes in the Golgi stack morphology and the transitional ER (tER) organization. Using conventional and immunoelectron microscopy and confocal immunofluorescence microscopy, we show that Golgi stacks were converted into clusters of vesicles and tubules, and that the tERs (marked by Sec23p) lost their focused organization and were now dispersed throughout the cytoplasm. However, we found that this morphologically altered exocytic pathway was nevertheless largely competent in anterograde protein transport using two different assays. The effects were specific for dp115. Depletion of the Drosophila homologues of GM130 and syntaxin 5 (dSed5p) did not lead to an effect on the tER organization, though the Golgi stacks were greatly vesiculated in the cells depleted of dSed5p. Taken together, these studies suggest that dp115 could be implicated in the architecture of both the Golgi stacks and the tER sites.

Show MeSH
Related in: MedlinePlus