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.
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21 July 2003
Article|
July 21 2003
A novel role for dp115 in the organization of tER sites in Drosophila
Vangelis Kondylis,
Vangelis Kondylis
1The Wellcome Trust Centre for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, Edinburgh, UK
2Department of Cell Biology, University Medical Centre Utrecht, Academic Ziekenhuis Utrecht, 3584CX Utrecht, Netherlands
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Catherine Rabouille
Catherine Rabouille
1The Wellcome Trust Centre for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, Edinburgh, UK
2Department of Cell Biology, University Medical Centre Utrecht, Academic Ziekenhuis Utrecht, 3584CX Utrecht, Netherlands
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Vangelis Kondylis
1The Wellcome Trust Centre for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, Edinburgh, UK
2Department of Cell Biology, University Medical Centre Utrecht, Academic Ziekenhuis Utrecht, 3584CX Utrecht, Netherlands
Catherine Rabouille
1The Wellcome Trust Centre for Cell Biology, Institute for Cell and Molecular Biology, University of Edinburgh, Edinburgh, UK
2Department of Cell Biology, University Medical Centre Utrecht, Academic Ziekenhuis Utrecht, 3584CX Utrecht, Netherlands
Address correspondence to Catherine Rabouille, Department of Cell Biology, University Medical Centre Utrecht, AZU Room G02.525, Heidelberglaan 100, 3584CX Utrecht, Netherlands. Tel.: 31-30-250 9280. Fax: 31-30-254 1797. E-mail: [email protected]
The online version of this article includes supplemental material.
*
Abbreviations used in this paper: GA, glutaraldehyde; IEM, immunoelectron microscopy; RNAi, RNA interference; tER, transitional ER.
Received:
January 30 2003
Revision Received:
May 15 2003
Accepted:
June 03 2003
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2003
J Cell Biol (2003) 162 (2): 185–198.
Article history
Received:
January 30 2003
Revision Received:
May 15 2003
Accepted:
June 03 2003
Citation
Vangelis Kondylis, Catherine Rabouille; A novel role for dp115 in the organization of tER sites in Drosophila . J Cell Biol 21 July 2003; 162 (2): 185–198. doi: https://doi.org/10.1083/jcb.200301136
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