Cog3p depletion blocks vesicle-mediated Golgi retrograde trafficking in HeLa cells.
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.
Zolov
Sergey N
SN
Department of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Lupashin
Vladimir V
VV
D033942:Adaptor Proteins, Vesicular Transport / Q000172:deficiency* / Q000378:metabolism; D000818:Animals; D020755:Coat Protein Complex I / Q000378:metabolism; D022162:Cytoplasmic Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D010750:Phosphoproteins / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; D022621:Shiga Toxin / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism
D033942:Adaptor Proteins, Vesicular Transport; C401396:COG3 protein, S cerevisiae; C498528:COG3 protein, human; D020755:Coat Protein Complex I; C107340:GOLIM4 protein, human; D008562:Membrane Glycoproteins; D010750:Phosphoproteins; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins; D022621:Shiga Toxin
J Cell Sci. 1994 Mar;107 ( Pt 3):529-37
8006071
J Cell Biol. 1994 Jul;126(1):53-64
8027186
Traffic. 2001 May;2(5):336-44
11350629
J Cell Biol. 1994 Nov;127(3):679-91
7962052
J Cell Biol. 1995 Dec;131(6 Pt 2):1715-26
8557739
J Cell Biol. 1996 Mar;132(6):985-98
8601597
Cell. 1997 Apr 4;89(1):149-58
9094723
Mol Biol Cell. 1997 Jun;8(6):1073-87
9201717
J Biol Chem. 1997 Aug 1;272(31):19554-61
9235960
Cell. 1997 Jul 25;90(2):335-49
9244307
J Cell Biol. 1998 Feb 9;140(3):541-51
9456315
Mol Biol Cell. 1998 Mar;9(3):623-35
9487131
J Cell Biol. 1998 Jun 1;141(5):1107-19
9606204
J Cell Biol. 1998 Jun 29;141(7):1489-502
9647643
J Cell Biol. 1998 Oct 19;143(2):319-31
9786945
J Biol Chem. 1998 Nov 6;273(45):29565-76
9792665
J Cell Biol. 1998 Nov 16;143(4):973-90
9817755
J Cell Biol. 1998 Dec 14;143(6):1505-21
9852147
Proc Natl Acad Sci U S A. 1999 Feb 2;96(3):915-20
9927668
EMBO J. 1999 Sep 15;18(18):4935-48
10487746
Nature. 2001 May 24;411(6836):494-8
11373684
J Biol Chem. 2001 Jun 22;276(25):22810-8
11292827
Biochem J. 2001 Aug 15;358(Pt 1):33-40
11485549
Biochimie. 2001 Aug;83(8):763-73
11530209
Mol Biol Cell. 2001 Oct;12(10):3152-60
11598199
J Cell Biol. 2001 Oct 15;155(2):227-38
11591729
J Cell Biol. 2001 Nov 12;155(4):543-55
11696556
Dev Cell. 2001 Oct;1(4):527-37
11703943
J Cell Biol. 2001 Dec 24;155(7):1199-212
11748249
J Cell Biol. 2001 Dec 24;155(7):1099-101
11756463
J Cell Biol. 2002 Apr 29;157(3):405-15
11980916
Mol Biol Cell. 2002 May;13(5):1484-500
12006647
J Cell Biol. 2002 May 13;157(4):631-43
12011112
J Biol Chem. 2002 Jun 14;277(24):21955-61
11929878
J Cell Sci. 2002 Jul 1;115(Pt 13):2627-37
12077354
Traffic. 2002 Sep;3(9):641-53
12191016
Annu Rev Cell Dev Biol. 2002;18:379-420
12142281
Mol Biol Cell. 2002 Oct;13(10):3493-507
12388752
Mol Biol Cell. 2003 Jan;14(1):190-200
12529436
Mol Membr Biol. 2003 Jul-Sep;20(3):197-207
12893528
Proc Natl Acad Sci U S A. 2004 Feb 3;101(5):1253-6
14736916
Mol Biol Cell. 2004 Apr;15(4):1506-18
14742712
Mol Biol Cell. 2004 May;15(5):2423-35
15004235
Nat Med. 2004 May;10(5):518-23
15107842
J Biol Chem. 2004 Jun 4;279(23):24640-8
15047703
Mol Biol Cell. 2004 Sep;15(9):4011-22
15215310
J Biol Chem. 2004 Sep 17;279(38):39814-23
15229219
J Cell Biol. 1986 May;102(5):1576-85
3700466
Cell. 1987 Aug 14;50(4):523-34
3038335
J Lipid Res. 1991 Mar;32(3):529-43
2066680
Mol Biol Cell. 1993 Jul;4(7):679-93
7691276
J Cell Biol. 1999 Nov 15;147(4):729-42
10562277