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Infection and Immunity, January 2001, p. 405-412, Vol. 69, No. 1
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.1.405-412.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Secreted Aspartic Proteinase Family of Candida tropicalis

Christophe Zaugg,1 Margarete Borg-von Zepelin,2 Utz Reichard,2 Dominique Sanglard,3 and Michel Monod1,*

Service de Dermatologie1 and Institut de Microbiologie,3 Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland, and Department of Bacteriology, University Clinics, University of Göttingen, Germany2

Received 21 August 2000/Returned for modification 2 October 2000/Accepted 18 October 2000

Medically important yeasts of the genus Candida secrete aspartic proteinases (Saps), which are of particular interest as virulence factors. Like Candida albicans, Candida tropicalis secretes in vitro one dominant Sap (Sapt1p) in a medium containing bovine serum albumin (BSA) as the sole source of nitrogen. Using the gene SAPT1 as a probe and under low-stringency hybridization conditions, three new closely related gene sequences, SAPT2 to SAPT4, encoding secreted proteinases were cloned from a C. tropicalis lambda EMBL3 genomic library. All bands identified by Southern blotting of EcoRI-digested C. tropicalis genomic DNA with SAPT1 could be assigned to a specific SAP gene. Therefore, the SAPT gene family of C. tropicalis is likely to contain only four members. Interestingly, the SAPT2 and SAPT3 gene products, Sapt2p and Sapt3p, which have not yet been detected in C. tropicalis cultures in vitro, were produced as active recombinant enzymes with the methylotrophic yeast Pichia pastoris as an expression system. As expected, reverse transcriptase PCR experiments revealed a strong SAPT1 signal with RNA extracted from cells grown in BSA medium. However, a weak signal was obtained with all other SAPT genes under several conditions tested, showing that these SAPT genes could be expressed at a basic level. Together, these experiments suggest that the gene products Sapt2p, Sapt3p, and Sapt4p could be produced under conditions yet to be described in vitro or during infection.

* Corresponding author. Mailing address: Service de Dermatologie, Laboratoire de Mycologie, BT422, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne, Switzerland. Phone: 41 21 314 0376. Fax: 41 21 314 0378. E-mail: Michel.Monod{at}chuv.hospvd.ch.

Infection and Immunity, January 2001, p. 405-412, Vol. 69, No. 1
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.1.405-412.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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