This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Osherov, N. Articles by Kontoyiannis, D. P. Search for Related Content PubMed PubMed Citation Articles by Osherov, N. Articles by Kontoyiannis, D. P.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, August 2002, p. 2462-2469, Vol. 46, No. 8
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.8.2462-2469.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Overexpression of Sbe2p, a Golgi Protein, Results in Resistance to Caspofungin in Saccharomyces cerevisiae

Department of Infectious Diseases, Infection Control, and Employee Health,² Department of Laboratory Medicine, The University of Texas M.D. Anderson Cancer Center, Houston, Texas¹

Received 20 September 2001/ Returned for modification 11 December 2001/ Accepted 26 April 2002

Caspofungin inhibits the synthesis of 1, 3-ß-D-glucan, an essential cell wall target in fungi. Genetic studies in the model yeast Saccharomyces cerevisiae have shown that mutations in FKS1 and FKS2 genes result in caspofungin resistance. However, direct demonstration of the role of gene overexpression in caspofungin resistance has been lacking. We transformed wild-type S. cerevisiae with an S. cerevisiae URA3-based GAL1 cDNA library and selected transformants in glucose synthetic complete plates lacking uracil (glucose SC minus uracil plates). We then moved the transformants to galactose SC minus uracil plates containing caspofungin (1 µg/ml) and looked for caspofungin-resistant colonies. We retested the candidates (true positives were sensitive on glucose caspofungin and resistant on galactose caspofungin media, respectively). We identified 16 caspofungin-resistant candidates. Restriction analysis and hybridization confirmed that 15 of the 16 clones were identical. We sequenced one of the cDNA clones and found that it contained the cDNA for SBE2. SBE2 has been described in S. cerevisiae to encode a Golgi protein involved in the transport of cell wall components (B. Santos and M. Snyder, Mol. Biol. Cell, 11:435-452, 2000). The SBE2 cDNA plasmid conferred again galactose-dependent caspofungin resistance when transformed back into the wild-type S. cerevisiae. Finally, the SBE2 deletion mutant was hypersensitive to caspofungin. In conclusion, overexpression of Sbe2p under the regulated control of the GAL1 promoter results in caspofungin resistance in S. cerevisiae. This transport pathway may provide insight into the tolerance or lack of sensitivity to caspofungin of some pathogenic fungi.

This article has been cited by other articles:

• Cota, J. M., Grabinski, J. L., Talbert, R. L., Burgess, D. S., Rogers, P. D., Edlind, T. D., Wiederhold, N. P. (2008). Increases in SLT2 Expression and Chitin Content Are Associated with Incomplete Killing of Candida glabrata by Caspofungin. Antimicrob. Agents Chemother. 52: 1144-1146 [Abstract] [Full Text]  
• Morris, M. I., Villmann, M. (2006). Echinocandins in the management of invasive fungal infections, part 1.. Am J Health Syst Pharm 63: 1693-1703 [Abstract] [Full Text]  
• Wiederhold, N. P., Kontoyiannis, D. P., Prince, R. A., Lewis, R. E. (2005). Attenuation of the Activity of Caspofungin at High Concentrations against Candida albicans: Possible Role of Cell Wall Integrity and Calcineurin Pathways. Antimicrob. Agents Chemother. 49: 5146-5148 [Abstract] [Full Text]  
• Krysan, D. J., Ting, E. L., Abeijon, C., Kroos, L., Fuller, R. S. (2005). Yapsins Are a Family of Aspartyl Proteases Required for Cell Wall Integrity in Saccharomyces cerevisiae. Eukaryot Cell 4: 1364-1374 [Abstract] [Full Text]  
• Markovich, S., Yekutiel, A., Shalit, I., Shadkchan, Y., Osherov, N. (2004). Genomic Approach to Identification of Mutations Affecting Caspofungin Susceptibility in Saccharomyces cerevisiae. Antimicrob. Agents Chemother. 48: 3871-3876 [Abstract] [Full Text]  
• Liu, W., May, G. S., Lionakis, M. S., Lewis, R. E., Kontoyiannis, D. P. (2004). Extra Copies of the Aspergillus fumigatus Squalene Epoxidase Gene Confer Resistance to Terbinafine: Genetic Approach to Studying Gene Dose-Dependent Resistance to Antifungals in A. fumigatus. Antimicrob. Agents Chemother. 48: 2490-2496 [Abstract] [Full Text]  
• Imhof, A., Balajee, S. A., Marr, K. A. (2003). New Methods To Assess Susceptibilities of Aspergillus Isolates to Caspofungin. J. Clin. Microbiol. 41: 5683-5688 [Abstract] [Full Text]  
• Agarwal, A. K., Rogers, P. D., Baerson, S. R., Jacob, M. R., Barker, K. S., Cleary, J. D., Walker, L. A., Nagle, D. G., Clark, A. M. (2003). Genome-wide Expression Profiling of the Response to Polyene, Pyrimidine, Azole, and Echinocandin Antifungal Agents in Saccharomyces cerevisiae. J. Biol. Chem. 278: 34998-35015 [Abstract] [Full Text]