The Journal of Experimental Medicine
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Published online 6 March 2000.
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© The Rockefeller University Press, 0022-1007/2000/3/871/ $5.00
The Journal of Experimental Medicine, Volume 191, Number 5, March 6, 2000 871-882

Original Article

Cryptococcus neoformans STE12{alpha} Regulates Virulence but Is Not Essential for Mating

Y.C. Changa, B.L. Wickesc, G.F. Millerb, L.A. Penoyera, and K.J. Kwon-Chunga

a Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, Office of the Director, National Institutes of Health, Bethesda, Maryland 20892
b Veterinary Resources Program, Office of the Director, National Institutes of Health, Bethesda, Maryland 20892
c Department of Microbiology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78284
LCI, NIAID, Bldg. 10, Rm. 11C304, National Institutes of Health, Bethesda, MD 20892.301-402-1003301-496-1602

June_Kwon-Chung{at}nih.gov

The Cryptococcus neoformans STE12{alpha} gene, a homologue of Saccharomyces cerevisiae STE12, exists only in mating type (MAT){alpha} cells. In S. cerevisiae, STE12 was required for mating and filament formation. In C. neoformans, haploid fruiting on filament agar required STE12{alpha}. The ability to form hyphae, however, was not affected by deletion of STE12{alpha} when convergently growing MATa strains were present. Furthermore, ste12{alpha} disruptants were fertile when mated with MATa strains, albeit with reduced mating frequency. Most importantly, the virulence of a ste12{alpha} disruptant of serotype D strain was significantly reduced in a mouse model. When the ste12{alpha} locus was reconstituted with the wild-type allele by cotransformation, virulence was restored. Histopathological analysis demonstrated a reduction in capsular size of yeast cells, less severe cystic lesions, and stronger immune responses in meninges of mice infected with ste12{alpha} cells than those of mice infected with STE12{alpha} cells. Using reporter gene constructs, we found that STE12{alpha} controls the expression of several phenotypes known to be involved in virulence, such as capsule and melanin production. These results demonstrate a clear molecular link between mating type and virulence in C. neoformans.

Key Words: haploid fruiting • mating assay • STE12 • cotransformation • virulence factor

Abbreviations used in this paper: GUS, β-glucuronidase; MAP, mitogen-activated protein; SLAD, synthetic low ammonia dextrose; YEPD, yeast extract peptone dextrose.

© 2000 government


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