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Journal of Bacteriology, April 2001, p. 2273-2279, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2273-2279.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Glucan Synthase Complex of Aspergillus fumigatus

A. Beauvais,1,* J. M. Bruneau,2 P. C. Mol,1,dagger M. J. Buitrago,1 R. Legrand,3 and J. P. Latgé1

Unité des Aspergillus, Institut Pasteur, Paris,1 and Infectious Disease Group, Biochemistry Department2 and Central Research Functions, Biophysics Department,3 Hoechst Marion Roussel, Romainville, France

Received 26 October 2000/Accepted 15 January 2001

The glucan synthase complex of the human pathogenic mold Aspergillus fumigatus has been investigated. The genes encoding the putative catalytic subunit Fks1p and four Rho proteins of A. fumigatus were cloned and sequenced. Sequence analysis showed that AfFks1p was a transmembrane protein very similar to other Fksp proteins in yeasts and in Aspergillus nidulans. Heterologous expression of the conserved internal hydrophilic domain of AfFks1p was achieved in Escherichia coli. Anti-Fks1p antibodies labeled the apex of the germ tube, as did aniline blue fluorochrome, which was specific for beta (1-3) glucans, showing that AfFks1p colocalized with the newly synthesized beta (1-3) glucans. AfRHO1, the most homologous gene to RHO1 of Saccharomyces cerevisiae, was studied for the first time in a filamentous fungus. AfRho proteins have GTP binding and hydrolysis consensus sequences identical to those of yeast Rho proteins and have a slightly modified geranylation site in AfRho1p and AfRho3p. Purification of the glucan synthase complex by product entrapment led to the enrichment of four proteins: Fks1p, Rho1p, a 100-kDa protein homologous to a membrane H+-ATPase, and a 160-kDa protein which was labeled by an anti-beta (1-3) glucan antibody and was homologous to ABC bacterial beta (1-2) glucan transporters.

* Corresponding author. Mailing address: Unité des Aspergillus, Institut Pasteur, 25 rue du docteur Roux, 75015 Paris, France. Phone: (33) 0145688225. Fax: (33) 0140613419. E-mail: abeauvai{at}pasteur.fr.

dagger Present address: Swammerdam Institute for Life Science, University of Amsterdam, Amsterdam, The Netherlands.

Journal of Bacteriology, April 2001, p. 2273-2279, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2273-2279.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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