The conserved oligomeric Golgi (COG) complex is an evolutionarily conserved multi-subunit protein complex that regulates membrane trafficking in eukaryotic cells. In this work we used short interfering RNA strategy to achieve an efficient knockdown (KD) of Cog3p in HeLa cells. For the first time, we have demonstrated that Cog3p depletion is accompanied by reduction in Cog1, 2, and 4 protein levels and by accumulation of COG complex-dependent (CCD) vesicles carrying v-SNAREs GS15 and GS28 and cis-Golgi glycoprotein GPP130. Some of these CCD vesicles appeared to be vesicular coat complex I (COPI) coated. A prolonged block in CCD vesicles tethering is accompanied by extensive fragmentation of the Golgi ribbon. Fragmented Golgi membranes maintained their juxtanuclear localization, cisternal organization and are competent for the anterograde trafficking of vesicular stomatitis virus G protein to the plasma membrane. In a contrast, Cog3p KD resulted in inhibition of retrograde trafficking of the Shiga toxin. Furthermore, the mammalian COG complex physically interacts with GS28 and COPI and specifically binds to isolated CCD vesicles.
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28 February 2005
Article|
February 22 2005
Cog3p depletion blocks vesicle-mediated Golgi retrograde trafficking in HeLa cells
Sergey N. Zolov,
Sergey N. Zolov
Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR 72205
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Vladimir V. Lupashin
Vladimir V. Lupashin
Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR 72205
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Sergey N. Zolov
Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR 72205
Vladimir V. Lupashin
Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR 72205
Correspondence to Vladimir Lupashin: [email protected]
Abbreviations used in this paper: CCD, COG complex-dependent; COG, conserved oligomeric Golgi; COPI, vesicular coat complex I; GalNAc-T2, N-acetylgalactosaminyltransferase-2; GalT-GFP, GFP-tagged β 1,4-galactosyltransferase; IF, immunofluorescence; IP, immunoprecipitation; KD, knockdown; PDI, protein disulphide isomerase; PNS, post-nuclear supernatant; siRNA, short interfering RNA; VSVG, vesicular stomatitis virus G protein; WB, Western blot.
Received:
December 01 2004
Accepted:
January 11 2005
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2005
J Cell Biol (2005) 168 (5): 747–759.
Article history
Received:
December 01 2004
Accepted:
January 11 2005
Citation
Sergey N. Zolov, Vladimir V. Lupashin; Cog3p depletion blocks vesicle-mediated Golgi retrograde trafficking in HeLa cells . J Cell Biol 28 February 2005; 168 (5): 747–759. doi: https://doi.org/10.1083/jcb.200412003
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