Yeast Genetics and Molecular Biology 1998
College Park, Maryland
August 1998


Name: Klis, Frans M.
Mailing Address: Inst Mol Cell Biol, BioCentrum Amsterdam, Kruislaan 318, Amsterdam, 1098 SM, The Netherlands
Email Address: klis@bio.uva.nl
Phone and Fax numbers: +31-20-525 7834, +31-20-525 7934

035

Evidence that beta-1,6-glucan synthesis takes place at the cell surface.


Roy C. Montijn (1), Edwin Vink (2), Wally Muller (3), Arie Verklei (3), Christiane Volland (4), Karin Seron (4), Herman van den Ende (2), Frans M. Klis (2)
(1) TNO Nutrition, Zeist, The Netherlands; (2) Inst Mol Cell Biol, BioCentrum Amsterdam, Kruislaan 318, Amsterdam, 1098 SM, The Netherlands; (3) Dept Mol Cell Biol, Univ Utrecht, The Netherlands; (4) Inst J Monod, Univ Paris VII, France

It has been proposed that beta-1,6-glucan is synthesized intracellularly in a stepwise process involving so-called KRE genes located in the various compartments of the secretory pathway. Here we propose that beta-1,6-glucan synthesis takes place at the cell surface. First, the proposed glucosyl transferases Kre6 and Skn1 are required for full N -glycosylation of the vacuolar protein carboxypeptidase Y, the plasma membrane-bound protein Gas1, the periplasmic enzyme invertase, and the cell wall protein alpha-agglutinin. This excludes that Kre6 and Skn1 are directly involved in beta-1,6-glucan synthesis. Second, using immunoelectron microscopy we find beta-1,6-glucan only at the cell surface, but never intracellularly. Third, plasma membrane-derived vesicles isolated by gel filtration contain beta-1,6-glucan in contrast to post-Golgi secretory vesicles. These data are consistent with a model in which KRE genes are involved in the synthesis of an acceptor structure for later beta-1,6-glucan addition and in which beta-1,6-glucan is synthesized at the plasma membrane.


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