Defective GM3 Synthesis in Cog2 Null Mutant CHO Cells Associates to Mislocalization of Lactosylceramide Sialyltransferase in the Golgi Complex

Overview

Journal Neurochem Res

Specialties Chemistry
Neurology

Date 2010 Nov 17

PMID 21080064

Citations 8

Authors

Waldo Spessott

Andrea Uliana

Hugo J F Maccioni

Affiliations

Soon will be listed here.

Abstract

The conserved oligomeric Golgi (COG) complex is a eight subunit (COG1 to 8) tethering complex involved in the retrograde trafficking of multiple Golgi processing proteins. Here we studied the glycolipid synthesis status in ldlC cells, a Cog2 null mutant CHO cell line. Biochemical studies revealed a block in the coupling between LacCer and GM3 synthesis, resulting in decreased levels of GM3 in these cells. Uncoupling was not attributable to decreased activity of the glycosyltransferase that uses LacCer as acceptor substrate (SialT1). Rather, immunocytochemical experiments evidenced a mislocalization of SialT1 as consequence of the lack of Cog2 in these cells. Co-immunoprecipitation experiments disclose a Cog2 mediated interaction of SialT1 with the COG complex member Cog1. Results indicate that cycling of some Golgi glycolipid glycosyltransferases depends on the participation of the COG complex and that deficiencies in COG complex subunits, by altering their traffic and localization, affect glycolipid composition.

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References

Wu X, Steet R, Bohorov O, Bakker J, Newell J, Krieger M . Mutation of the COG complex subunit gene COG7 causes a lethal congenital disorder. Nat Med. 2004; 10(5):518-23. DOI: 10.1038/nm1041. View

Vasile E, Oka T, Ericsson M, Nakamura N, Krieger M . IntraGolgi distribution of the Conserved Oligomeric Golgi (COG) complex. Exp Cell Res. 2006; 312(16):3132-41. DOI: 10.1016/j.yexcr.2006.06.005. View

Kingsley D, Kozarsky K, Segal M, Krieger M . Three types of low density lipoprotein receptor-deficient mutant have pleiotropic defects in the synthesis of N-linked, O-linked, and lipid-linked carbohydrate chains. J Cell Biol. 1986; 102(5):1576-85. PMC: 2114220. DOI: 10.1083/jcb.102.5.1576. View

Ungar D, Oka T, Brittle E, Vasile E, Lupashin V, Chatterton J . Characterization of a mammalian Golgi-localized protein complex, COG, that is required for normal Golgi morphology and function. J Cell Biol. 2002; 157(3):405-15. PMC: 2173297. DOI: 10.1083/jcb.200202016. View

Ungar D, Oka T, Vasile E, Krieger M, Hughson F . Subunit architecture of the conserved oligomeric Golgi complex. J Biol Chem. 2005; 280(38):32729-35. DOI: 10.1074/jbc.M504590200. View

Yu R, Macala L, Taki T, Weinfield H, Yu F . Developmental changes in ganglioside composition and synthesis in embryonic rat brain. J Neurochem. 1988; 50(6):1825-9. DOI: 10.1111/j.1471-4159.1988.tb02484.x. View

Todeschini A, Hakomori S . Functional role of glycosphingolipids and gangliosides in control of cell adhesion, motility, and growth, through glycosynaptic microdomains. Biochim Biophys Acta. 2007; 1780(3):421-33. PMC: 2312458. DOI: 10.1016/j.bbagen.2007.10.008. View

Panzetta P, Gravotta D, Maccioni H . Biosynthesis and expression of gangliosides during differentiation of chick embryo retina cells in vitro. J Neurochem. 1987; 49(6):1763-71. DOI: 10.1111/j.1471-4159.1987.tb02434.x. View

Krieger M, Brown M, Goldstein J . Isolation of Chinese hamster cell mutants defective in the receptor-mediated endocytosis of low density lipoprotein. J Mol Biol. 1981; 150(2):167-84. DOI: 10.1016/0022-2836(81)90447-2. View

10.

Maxzud M, Daniotti J, Maccioni H . Functional coupling of glycosyl transfer steps for synthesis of gangliosides in Golgi membranes from neural retina cells. J Biol Chem. 1995; 270(34):20207-14. DOI: 10.1074/jbc.270.34.20207. View

11.

Krieger M, Reddy P, Kozarsky K, Kingsley D, Hobbie L, Penman M . Analysis of the synthesis, intracellular sorting, and function of glycoproteins using a mammalian cell mutant with reversible glycosylation defects. Methods Cell Biol. 1989; 32:57-84. DOI: 10.1016/s0091-679x(08)61167-x. View

12.

Reynders E, Foulquier F, Leao Teles E, Quelhas D, Morelle W, Rabouille C . Golgi function and dysfunction in the first COG4-deficient CDG type II patient. Hum Mol Genet. 2009; 18(17):3244-56. PMC: 2722986. DOI: 10.1093/hmg/ddp262. View

13.

Maccioni H . Glycosylation of glycolipids in the Golgi complex. J Neurochem. 2007; 103 Suppl 1:81-90. DOI: 10.1111/j.1471-4159.2007.04717.x. View

14.

Podos S, Reddy P, Ashkenas J, Krieger M . LDLC encodes a brefeldin A-sensitive, peripheral Golgi protein required for normal Golgi function. J Cell Biol. 1994; 127(3):679-91. PMC: 2120235. DOI: 10.1083/jcb.127.3.679. View

15.

Whyte J, Munro S . Vesicle tethering complexes in membrane traffic. J Cell Sci. 2002; 115(Pt 13):2627-37. DOI: 10.1242/jcs.115.13.2627. View

16.

Lippincott-Schwartz J, Yuan L, Bonifacino J, Klausner R . Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER. Cell. 1989; 56(5):801-13. PMC: 7173269. DOI: 10.1016/0092-8674(89)90685-5. View

17.

Shestakova A, Zolov S, Lupashin V . COG complex-mediated recycling of Golgi glycosyltransferases is essential for normal protein glycosylation. Traffic. 2006; 7(2):191-204. DOI: 10.1111/j.1600-0854.2005.00376.x. View

18.

Chatterton J, Hirsch D, Schwartz J, Bickel P, Rosenberg R, Lodish H . Expression cloning of LDLB, a gene essential for normal Golgi function and assembly of the ldlCp complex. Proc Natl Acad Sci U S A. 1999; 96(3):915-20. PMC: 15325. DOI: 10.1073/pnas.96.3.915. View

19.

Rhee S, Starr T, Forsten-Williams K, Storrie B . The steady-state distribution of glycosyltransferases between the Golgi apparatus and the endoplasmic reticulum is approximately 90:10. Traffic. 2005; 6(11):978-90. DOI: 10.1111/j.1600-0854.2005.00333.x. View

20.

Quintero C, Giraudo C, Villarreal M, Montich G, Maccioni H . Identification of a site in Sar1 involved in the interaction with the cytoplasmic tail of glycolipid glycosyltransferases. J Biol Chem. 2010; 285(39):30340-6. PMC: 2943283. DOI: 10.1074/jbc.M110.128868. View