Characterization and regulation of the trehalose synthesis pathway and its importance in the pathogenicity of Cryptococcus neoformans.
The disaccharide trehalose has been found to play diverse roles, from energy source to stress protectant, and this sugar is found in organisms as diverse as bacteria, fungi, plants, and invertebrates but not in mammals. Recent studies in the pathobiology of Cryptococcus neoformans identified the presence of a functioning trehalose pathway during infection and suggested its importance for C. neoformans survival in the host. Therefore, in C. neoformans we created null mutants of the trehalose-6-phosphate (T6P) synthase (TPS1), trehalose-6-phophate phosphatase (TPS2), and neutral trehalase (NTH1) genes. We found that both TPS1 and TPS2 are required for high-temperature (37 degrees C) growth and glycolysis but that the block at TPS2 results in the apparent toxic accumulation of T6P, which makes this enzyme a fungicidal target. Sorbitol suppresses the growth defect in the tps1 and tps2 mutants at 37 degrees C, which supports the hypothesis that these sugars (trehalose and sorbitol) act primarily as stress protectants for proteins and membranes during exposure to high temperatures in C. neoformans. The essential nature of this pathway for disease was confirmed when a tps1 mutant strain was found to be avirulent in both rabbits and mice. Furthermore, in the system of the invertebrate C. elegans, in which high in vivo temperature is no longer an environmental factor, attenuation in virulence was still noted with the tps1 mutant, and this supports the hypothesis that the trehalose pathway in C. neoformans is involved in more host survival mechanisms than simply high-temperature stresses and glycolysis. These studies in C. neoformans and previous studies in other pathogenic fungi support the view of the trehalose pathway as a selective fungicidal target for use in antifungal development.
Petzold
Elizabeth Wills
EW
Department of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Himmelreich
Uwe
U
Mylonakis
Eleftherios
E
Rude
Thomas
T
Toffaletti
Dena
D
Cox
Gary M
GM
Miller
Jackie L
JL
Perfect
John R
JR
D000818:Animals; D003453:Cryptococcosis / Q000382:microbiology*; D003455:Cryptococcus neoformans / Q000201:enzymology / Q000235:genetics / Q000472:pathogenicity*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D006019:Glycolysis / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011817:Rabbits; D013012:Sorbitol / Q000494:pharmacology; D013403:Sugar Phosphates / Q000032:analysis / Q000378:metabolism; D013696:Temperature; D014198:Trehalase / Q000235:genetics / Q000378:metabolism; D014199:Trehalose / Q000031:analogs & derivatives / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D014774:Virulence / Q000235:genetics
D005656:Fungal Proteins; D013403:Sugar Phosphates; C082722:trehalose-6-phosphate; D013012:Sorbitol; D014199:Trehalose; D005964:Glucosyltransferases; C099692:trehalose synthase; C023180:trehalose-6-phosphate synthase; D014198:Trehalase