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J Biol Chem, Vol. 275, Issue 20, 14882-14889, May 19, 2000

Glycosylphosphatidylinositol-anchored Glucanosyltransferases Play an Active Role in the Biosynthesis of the Fungal Cell Wall^*

Isabelle Mouyna^§, Thierry Fontaine, Marina Vai^¶, Michel Monod^**, William A. Fonzi, Michel Diaquin, Laura Popolo^¶, Robbert P. Hartland, and Jean-Paul Latgé

From the  Institut Pasteur, Laboratoire des Aspergillus, 25 Rue du Docteur Roux, 75724 Paris, Cedex 15, France, ^¶ Università degli Studi di Milano, Dipartimento di Fisiologia, e Biochimica Generali, Via Celoria 26, 20133 Milan, Italy, ^** Laboratoire de Mycologie, Service de Dermatologie, Hopital de Beaumont, 04-423 CHUV 1011 Lausanne, Switzerland, and  Department of Microbiology and Immunology, School of Medicine, Georgetown University Medical Center, Washington, D. C. 20007

A novel 1,3--glucanosyltransferase isolated from the cell wall of Aspergillus fumigatus was recently characterized. This enzyme splits internally a 1,3--glucan molecule and transfers the newly generated reducing end to the non-reducing end of another 1,3--glucan molecule forming a 1,3- linkage, resulting in the elongation of 1,3--glucan chains. The GEL1 gene encoding this enzyme was cloned and sequenced. The predicted amino acid sequence of Gel1p was homologous to several yeast protein families encoded by GAS of Saccharomyces cerevisiae, PHR of Candida albicans, and EPD of Candida maltosa. Although the expression of these genes is required for correct morphogenesis in yeast, the biochemical function of the encoded proteins was unknown. The biochemical assays performed on purified recombinant Gas1p, Phr1p, and Phr2p showed that these proteins have a 1,3--glucanosyltransferase activity similar to that of Gel1p. Biochemical data and sequence analysis have shown that Gel1p is attached to the membrane through a glycosylphosphatidylinositol in a similar manner as the yeast homologous proteins. The activity has been also detected in membrane preparations, showing that this 1,3--glucanosyltransferase is indeed active in vivo. Our results show that transglycosidases anchored to the plasma membrane via glycosylphosphatidylinositols can play an active role in fungal cell wall synthesis.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EMBL Data Bank with accession number(s) AF072700.

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