Author:

Keywords:

protein-coupled receptor, yeast saccharomyces-cerevisiae, trehalose synthase complex, trehalose-6-phosphate phosphatase, decreases infectivity, stress resistance, hypha formation, growth, mutant, disruption, Science & Technology, Life Sciences & Biomedicine, Immunology, Infectious Diseases, PROTEIN-COUPLED RECEPTOR, YEAST SACCHAROMYCES-CEREVISIAE, TREHALOSE SYNTHASE COMPLEX, TREHALOSE-6-PHOSPHATE PHOSPHATASE, DECREASES INFECTIVITY, STRESS RESISTANCE, HYPHA FORMATION, GROWTH, MUTANT, DISRUPTION, Animals, Candida albicans, Candidiasis, Fungal Proteins, Gene Deletion, Gene Expression Regulation, Fungal, Genes, Fungal, Glucosyltransferases, Macrophages, Peritoneal, Mice, Mutation, Receptors, G-Protein-Coupled, Sugar Phosphates, Trehalose, Virulence, 06 Biological Sciences, 07 Agricultural and Veterinary Sciences, 11 Medical and Health Sciences, Microbiology, 3107 Microbiology, 3204 Immunology, 3207 Medical microbiology

Abstract:

Inhibition of the biosynthesis of trehalose, a well-known stress protectant in pathogens, is an interesting approach for antifungal or antibacterial therapy. Deletion of TPS2, encoding trehalose-6-phosphate (T6P) phosphatase, results in strongly reduced virulence of Candida albicans due to accumulation of TO instead of trehalose in response to stress. To further aggravate the deregulation in the pathogen, we have additionally deleted the GPR1 gene, encoding the nutrient receptor that activates the cyclic AMP-protein kinase A signaling pathway, which negatively regulates trehalose accumulation in yeasts. A gpr1 mutant is strongly affected in morphogenesis on solid media as well as in vivo in a mouse model but has only a slightly decreased virulence. The gpr1 tps2 double mutant, on the other hand, is completely avirulent in a mouse model for systemic infection. This strain accumulates very high UP levels under stress conditions and has a growth defect at higher temperatures. We also show that a tps2 mutant is more sensitive to being killed by macrophages than the wild type or the gpr1 mutant. A double mutant has susceptibility similar to that of the single tps2 mutant. For morphogenesis on solid media, on the other hand, the gpr1 tps2 mutant shows a phenotype similar to that of the single gpr1 mutant. Taken together these results show that there is synergism between Gpr1 and Tps2 and that their combined inactivation results in complete avirulence. Combination therapy targeting both proteins may prove highly effective against pathogenic fungi with increased resistance to the currently used antifungal drugs.