Diploid strains of the pathogenic basidiomycete Cryptococcus neoformans are thermally dimorphic.
Cryptococcus neoformans is an opportunistic human pathogenic fungus with a defined sexual cycle. Clinical and environmental isolates of C. neoformans are haploid, and the diploid stage of the lifecycle is thought to be transient and unstable. In contrast, we find that diploid strains are readily obtained following genetic crosses of congenic MATalpha and MATa strains. At 37 degrees C, the diploid strains grow as yeast cells with a single nucleus that is larger than a haploid nucleus, contains a 2n content of DNA by FACS analysis, and is heterozygous for the MATalpha and MATa loci. At 24 degrees C, these diploid self-fertile strains filament and sporulate, producing recombinant haploid progeny in which meiotic segregation has occurred. In contrast to dikaryotic filament cells that are typically linked by fused clamp connections during mating, self-fertile diploid strains produce monokaryotic filament cells with unfused clamp connections. We also show that these diploid strains can be transformed and sporulated and that an integrated selectable marker segregates in a mendelian fashion. The diploid state could play novel roles in the lifecycle and virulence of the organism and can be exploited for the analysis of essential genes. Finally, the observation that dimorphism is thermally regulated suggests similarities between the lifecycle of C. neoformans and other thermally dimorphic human pathogenic fungi, including Histoplasma capsulatum, Blastomyces dermatitidis, Coccidioides immitis, Paracoccidioides brasiliensis, and Sporothrix schenkii.
Sia
R A
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Department of Genetics, Duke University Medical Center, Durham, North Carolina, 27710, USA.
Lengeler
K B
KB
Heitman
J
J
D003433:Crosses, Genetic; D003455:Cryptococcus neoformans / Q000166:cytology / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D004271:DNA, Fungal / Q000032:analysis; D004171:Diploidy; D005434:Flow Cytometry; D015966:Gene Expression Regulation, Fungal; D006238:Haploidy; D006579:Heterozygote; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D008856:Microscopy, Fluorescence; D011995:Recombination, Genetic; D012097:Repressor Proteins / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D013172:Spores, Fungal / Q000235:genetics / Q000502:physiology; D013696:Temperature; D014170:Transformation, Genetic; D014774:Virulence
D004271:DNA, Fungal; D018398:Homeodomain Proteins; C096063:MATA1 protein, S cerevisiae; C062928:MATA2 protein, S cerevisiae; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins