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

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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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Acknowledgments

This work was supported in part by Grants from Secretaría de Ciencia y Tecnología, Universidad Nacional de Córdoba, Argentina, Agencia Nacional de Promoción Científica y Tecnológica, Argentina and Ministerio de Ciencia y Tecnología de la Provincia de Córdoba. The technical assistance of G. Schachner and S. Deza with cell cultures and of M. C. Sampedro with confocal microscopy is also acknowledged. H. J. F. M. is a Career Investigator of CONICET (Argentina).

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  1. Centro de Investigaciones en Química Biológica de Córdoba, CIQUIBIC (UNC-CONICET), Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Haya de la Torre s/n—Pabellón Argentina Ala Oeste, 5000, Córdoba, Argentina

    Waldo Spessott, Andrea Uliana & Hugo J. F. Maccioni

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  1. Waldo Spessott
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  2. Andrea Uliana
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  3. Hugo J. F. Maccioni
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Correspondence to Hugo J. F. Maccioni.

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This work is dedicated to Dr. Abel L. Lajtha on occasion of his retirement as Editor-in-Chief of Neurochemical Research, of which he was founder and which he continuously improved during the last 35 years. In addition to his solid scientific work and his decisive contribution to the development of Neurochemistry, his kind personality has made very pleasant the time I spent as Member of the Editorial Board.

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Spessott, W., Uliana, A. & Maccioni, H.J.F. Defective GM3 Synthesis in Cog2 Null Mutant CHO Cells Associates to Mislocalization of Lactosylceramide Sialyltransferase in the Golgi Complex. Neurochem Res 35, 2161–2167 (2010). https://doi.org/10.1007/s11064-010-0319-8

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