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Eukaryotic Cell, June 2002, p. 432-439, Vol. 1, No. 3
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.3.432-439.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of CPR{alpha}, a MAT{alpha}-Specific Pheromone Receptor Gene of Cryptococcus neoformans

Seyung Chung,1 Marvin Karos,1,{dagger} Yun C. Chang,1 Jan Lukszo,2 Brian L. Wickes,3 and Kyung J. Kwon-Chung1*

Molecular Microbiology Section, Laboratory of Clinical Investigation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892,1 Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland 20852,2 Department of Microbiology, University of Texas, Health Center at San Antonio, San Antonio, Texas 782843

Received 4 December 2001/ Accepted 1 March 2002

The putative Cryptococcus neoformans pheromone receptor gene CPR{alpha} was isolated and studied for its role in mating and filamentation. CPR{alpha} is MAT{alpha} specific and located adjacent to STE12{alpha} at the MAT{alpha} locus. It encodes a protein which possesses high sequence similarity to the seven-transmembrane class of G-protein-coupled pheromone receptors reported for other basidiomycetous fungi. Strains containing a deletion of the CPR{alpha} gene exhibited drastic reductions in mating efficiency but were not completely sterile. {Delta}cpr{alpha} cells displayed wild-type mating efficiency when reconstituted with the wild-type CPR{alpha} gene. Hyphal production on filament agar was not affected in the {Delta}cpr{alpha} strain, indicating no significant role for CPR{alpha} in sensing environmental cues during haploid fruiting. The wild-type MAT{alpha} CPR{alpha} strain produced abundant hyphae in response to synthetic MATa pheromone; however, the hyphal response to pheromone by {Delta}cpr{alpha} cells was significantly reduced. Exposure of wild-type cells to synthetic MATa pheromone for 2 h induced MF{alpha} pheromone expression, whereas unexposed cells showed only basal levels of the MF{alpha} transcript. The {Delta}cpr{alpha} cells, however, exhibited only basal levels of MF{alpha} message with or without pheromone exposure, suggesting that CPR{alpha} and MF{alpha} are components of the same signaling pathway.

* Corresponding author. Mailing address: Bldg. 10, 11C304, NIH, Bethesda, MD 20892. Phone: (301) 496-1602. Fax: (301) 402-1003. E-mail: june_kwon-chung{at}nih.gov.

{dagger} Present address: BASF, Aktiengesellschaft, GVF-A30, 67056 Ludwigshafen, Germany.

Eukaryotic Cell, June 2002, p. 432-439, Vol. 1, No. 3
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.3.432-439.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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