title: Protein Composition of Infectious Spores Reveals Novel Sexual Development and Germination Factors in Cryptococcus.
abstract: Spores are an essential cell type required for long-term survival across diverse organisms in the tree of life and are a hallmark of fungal reproduction, persistence, and dispersal. Among human fungal pathogens, spores are presumed infectious particles, but relatively little is known about this robust cell type. Here we used the meningitis-causing fungus Cryptococcus neoformans to determine the roles of spore-resident proteins in spore biology. Using highly sensitive nanoscale liquid chromatography/mass spectrometry, we compared the proteomes of spores and vegetative cells (yeast) and identified eighteen proteins specifically enriched in spores. The genes encoding these proteins were deleted, and the resulting strains were evaluated for discernable phenotypes. We hypothesized that spore-enriched proteins would be preferentially involved in spore-specific processes such as dormancy, stress resistance, and germination. Surprisingly, however, the majority of the mutants harbored defects in sexual development, the process by which spores are formed. One mutant in the cohort was defective in the spore-specific process of germination, showing a delay specifically in the initiation of vegetative growth. Thus, by using this in-depth proteomics approach as a screening tool for cell type-specific proteins and combining it with molecular genetics, we successfully identified the first germination factor in C. neoformans. We also identified numerous proteins with previously unknown functions in both sexual development and spore composition. Our findings provide the first insights into the basic protein components of infectious spores and reveal unexpected molecular connections between infectious particle production and spore composition in a pathogenic eukaryote.
authors: Department of Biomolecular Chemistry, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors: Mingwei
authors: M
authors: Huang
authors: Department of Biomolecular Chemistry, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors: Alexander S
authors: AS
authors: Hebert
authors: Department of Biomolecular Chemistry, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America; Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America; Genome Center of Wisconsin, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors: Joshua J
authors: JJ
authors: Coon
authors: Department of Biomolecular Chemistry, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America; Department of Medical Microbiology & Immunology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
authors: Christina M
authors: CM
authors: Hull
mesh_terms: D002853:Chromatography, Liquid; D003455:Cryptococcus neoformans / Q000235:genetics / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics*; D049770:Genes, Mating Type, Fungal / Q000235:genetics*; D013058:Mass Spectrometry; D020543:Proteome / Q000235:genetics; D013172:Spores, Fungal / Q000235:genetics* / Q000378:metabolism; D015003:Yeasts / Q000235:genetics* / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D020543:Proteome