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Eukaryotic Cell, February 2004, p. 40-51, Vol. 3, No. 1
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.1.40-51.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Wangiella (Exophiala) dermatitidis WdChs5p, a Class V Chitin Synthase, Is Essential for Sustained Cell Growth at Temperature of Infection

Hongbo Liu,1 Sarah Kauffman,2 Jeffrey M. Becker,2 and Paul J. Szaniszlo1*

Section of Molecular Genetics and Microbiology, School of Biological Science and Institute of Cellular and Molecular Biology, The University of Texas at Austin, Austin, Texas 78712,1 Microbiology Department, University of Tennessee, Knoxville, Tennessee 379192

Received 2 October 2003/ Accepted 4 December 2003

The chitin synthase structural gene WdCHS5 was isolated from the black fungal pathogen of humans Wangiella dermatitidis. Sequence analysis revealed that the gene has a myosin motor-like-encoding region at its 5' end and a chitin synthase (class V)-encoding region at its 3' end. Northern blotting showed that WdCHS5 is expressed at high levels under conditions of stress. Analysis of the 5' upstream region of WdCHS5 fused to a reporter gene indicated that one or more of the potential regulatory elements present may have contributed to the high expression levels. Disruption of WdCHS5 produced mutants that grow normally at 25°C but have severe growth and cellular abnormalities at 37°C. Osmotic stabilizers, such as sorbitol and sucrose, rescued the wild-type phenotype, which indicated that the loss of WdChs5p causes cell wall integrity defects. Animal survival tests with a mouse model of acute infection showed that all wdchs5{Delta} mutants are less virulent than the parental strain. Reintroduction of the WdCHS5 gene into the wdchs5{Delta} mutants abolished the temperature-sensitive phenotype and reestablished their virulence. We conclude that the product of WdCHS5 is required for the sustained growth of W. dermatitidis at 37°C and is of critical importance to its virulence.

* Corresponding author. Mailing address: Section of Molecular Genetics and Microbiology, University of Texas at Austin, Austin, TX 78712-1095. Phone: (512) 471-3384. Fax: (512) 471-7088. E-mail: pjszaniszlo{at}mail.utexas.edu.

Eukaryotic Cell, February 2004, p. 40-51, Vol. 3, No. 1
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.1.40-51.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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