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Eukaryotic Cell, April 2004, p. 385-392, Vol. 3, No. 2
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.2.385-392.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Cryptococcus neoformans CAP59 (or Cap59p) Is Involved in the Extracellular Trafficking of Capsular Glucuronoxylomannan

Javier García-Rivera,1 Yun C. Chang,2 K. J. Kwon-Chung,2 and Arturo Casadevall1,3*

Department of Microbiology and Immunology,1 Department of Medicine, Albert Einstein College of Medicine, Bronx, New York 10461,3 Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 208922

Received 17 July 2003/ Accepted 3 January 2004

Several genes are essential for Cryptococcus neoformans capsule synthesis, but their functions are unknown. We examined the localization of glucuronoxylomannan (GXM) in strain B-3501 and in cap59 mutants B-4131 and C536. Wild-type strain B-3501 showed a visible capsule by India ink staining and immunofluorescence with anticapsular monoclonal antibodies (MAbs) 12A1 and 18B7. B-4131, a mutant containing a missense mutation in CAP59, showed no capsule by India ink staining but revealed the presence of capsular polysaccharide on the cell surface by immunofluorescence. The cap59 gene deletion mutant (C536), however, did not show a capsule by either India ink staining or immunofluorescence. Analysis of cell lysates for GXM by enzyme-linked immunosorbent assay revealed GXM in C536 samples. Furthermore, the epitopes recognized by MAbs 12A1, 2D10, 13F1, and 18B7 were each detected in the cytoplasm of all strains by immunogold electron microscopy, although there were differences in location consistent with differences in epitope synthesis and/or transport. In addition, the cells of B-3501 and B-4131, but not those of the cap59 deletant, assimilated raffinose or urea. Hence, the missense mutation of CAP59 in B-4131 partially hampered the trafficking of GXM but allowed the secretion of enzymes involved in hydrolysis of raffinose or urea. Furthermore, the cell diameter and volume for strain C536 are higher than those for strain B-3501 or B-4131 and may suggest the accumulation of cellular material in the cytoplasm. Our results suggest that CAP59 is involved in capsule synthesis by participating in the process of GXM (polysaccharide) export.

* Corresponding author. Mailing address: Albert Einstein College of Medicine, Department of Medicine, 1300 Morris Park Ave., 610 Belfer, Bronx, NY 10461. Phone: (718) 430-3659. Fax: (718) 430-8701. E-mail: casadeva{at}aecom.yu.edu.

Eukaryotic Cell, April 2004, p. 385-392, Vol. 3, No. 2
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.2.385-392.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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