Localization of an alpha 1,2 galactosyltransferase activity to the Golgi apparatus of Schizosaccharomyces pombe

TG Chappell, MA Hajibagheri, K Ayscough, M Pierce and G Warren

Imperial Cancer Research Fund, London, United Kingdom.

We have cloned a gene encoding an alpha 1,2 galactosyltransferase activity from Schizosaccharomyces pombe. The open reading frame of the gene (gma12 for galactomannan, alpha 1,2), combined with the previous protein purification (Chappell and Warren, 1989), predicts an O-linked glycoprotein with type II transmembrane topology. By homologous gene disruption, we have demonstrated that the gma12 gene product (gma12p) is nonessential. The deletion strain (gma12-D10::ura4) has a significantly reduced level of galactosyltransferase activity relative to the parental strain, but both in situ lectin binding and in vitro biochemical assays demonstrate the presence of further galactosyltransferase activity in addition to gma12p. Although gma12p is not the only galactosyltransferase in S. pombe, it produces a unique carbohydrate structure on the surface of the yeast cells. We have generated a polyclonal antiserum against this carbohydrate epitope and shown that gma12p is capable of synthesizing the epitope both in vitro and in vivo. Electron microscopic localization of the gma12+ specific epitope in gma12+ cells revealed that gma12p synthesizes the carbohydrate structure in the Golgi apparatus, and subsequent intracellular transport distributes the epitope to later stages of the secretory pathway. The immunolocalization studies confirm the presence of one or more galactosyltransferase activities in the Golgi apparatus in fission yeast.

Volume 5, Issue 5, pp. 519-528, 05/01/1994
Copyright © 1994 by The American Society for Cell Biology

This article has been cited by other articles:

				P. Magnelli, J. F. Cipollo, D. M. Ratner, J. Cui, D. Kelleher, R. Gilmore, C. E. Costello, P. W. Robbins, and J. Samuelson Unique Asn-linked Oligosaccharides of the Human Pathogen Entamoeba histolytica J. Biol. Chem., June 27, 2008; 283(26): 18355 - 18364. [Abstract] [Full Text] [PDF]

				C. D'Alessio, J. J. Caramelo, and A. J. Parodi Absence of Nucleoside Diphosphatase Activities in the Yeast Secretory Pathway Does Not Abolish Nucleotide Sugar-dependent Protein Glycosylation J. Biol. Chem., December 9, 2005; 280(49): 40417 - 40427. [Abstract] [Full Text] [PDF]

				E. N. Andreishcheva, J. P. Kunkel, T. R. Gemmill, and R. B. Trimble Five Genes Involved in Biosynthesis of the Pyruvylated Gal{beta}1,3-Epitope in Schizosaccharomyces pombe N-Linked Glycans J. Biol. Chem., August 20, 2004; 279(34): 35644 - 35655. [Abstract] [Full Text] [PDF]

				M. Nakamura-Kubo, T. Nakamura, A. Hirata, and C. Shimoda The Fission Yeast spo14+ Gene Encoding a Functional Homologue of Budding Yeast Sec12 Is Required for the Development of Forespore Membranes Mol. Biol. Cell, March 1, 2003; 14(3): 1109 - 1124. [Abstract] [Full Text] [PDF]

				H. Abe, Y.-i. Shimma, and Y. Jigami In vitro oligosaccharide synthesis using intact yeast cells that display glycosyltransferases at the cell surface through cell wall-anchored protein Pir Glycobiology, February 1, 2003; 13(2): 87 - 95. [Abstract] [Full Text] [PDF]

				H. Cheng, R. Sugiura, W. Wu, M. Fujita, Y. Lu, S. O. Sio, R. Kawai, K. Takegawa, H. Shuntoh, and T. Kuno Role of the Rab GTP-Binding Protein Ypt3 in the Fission Yeast Exocytic Pathway and Its Connection to Calcineurin Function Mol. Biol. Cell, August 1, 2002; 13(8): 2963 - 2976. [Abstract] [Full Text] [PDF]

				J. Liu, X. Tang, H. Wang, S. Oliferenko, and M. K. Balasubramanian The Localization of the Integral Membrane Protein Cps1p to the Cell Division Site is Dependent on the Actomyosin Ring and the Septation-Inducing Network in Schizosaccharomyces pombe Mol. Biol. Cell, March 1, 2002; 13(3): 989 - 1000. [Abstract] [Full Text] [PDF]

				A. Feoktistova, P. Magnelli, C. Abeijon, P. Perez, R. L. Lester, R. C. Dickson, and K. L. Gould Coordination Between Fission Yeast Glucan Formation and Growth Requires a Sphingolipase Activity Genetics, August 1, 2001; 158(4): 1397 - 1411. [Abstract] [Full Text] [PDF]

				S. B. Southard, C. A. Specht, C. Mishra, J. Chen-Weiner, and P. W. Robbins Molecular Analysis of the Candida albicans Homolog of Saccharomyces cerevisiae MNN9, Required for Glycosylation of Cell Wall Mannoproteins J. Bacteriol., December 15, 1999; 181(24): 7439 - 7448. [Abstract] [Full Text]

				J. Jungmann, J. C. Rayner, and S. Munro The Saccharomyces cerevisiae Protein Mnn10p/Bed1p Is a Subunit of a Golgi Mannosyltransferase Complex J. Biol. Chem., March 5, 1999; 274(10): 6579 - 6585. [Abstract] [Full Text] [PDF]

				K. Shibayama, S. Ohsuka, T. Tanaka, Y. Arakawa, and M. Ohta Conserved Structural Regions Involved in the Catalytic Mechanism of Escherichia coli K-12 WaaO (RfaI) J. Bacteriol., October 15, 1998; 180(20): 5313 - 5318. [Abstract] [Full Text]

				S. K. Roy, T. Yoko-o, H. Ikenaga, and Y. Jigami Functional Evidence for UDP-galactose Transporter in Saccharomyces cerevisiae through the in Vivo Galactosylation and in Vitro Transport Assay J. Biol. Chem., January 30, 1998; 273(5): 2583 - 2590. [Abstract] [Full Text] [PDF]