IADR Abstract Archives
Identifying contact points for the development of next-generation antifungals
Objectives: Alanine substitutions were introduced by site-directed mutagenesis in three of these amino acid residues in Saccharomyces cerevisiae LDM (L147A, M313A and L480A) to assess their impact on LDM function and response to azole drugs.
Methods: Three DNA cassettes generated using PCR with mutagenic oligonucleotide primers were transformed into yeast host strains. Liquid microdilution susceptibility assays were used to determine the minimal inhibitory drug concentrations of the mutant strains.
Results: None of the mutations were lethal and they did not appear to deleteriously affect enzyme function. Instead, these mutations increased susceptibility to short- and medium-tailed, but not long-tailed azole antifungals. Alanine mutagenesis of the other four conserved LBP residues of interest (Y72, F241, H381, and S382) was previously found to give similar phenotypes.
Conclusions: This project analysed amino acids conserved within the LBP of LDM of 10 important fungal pathogens. Mutational analysis has helped us understand the impact these 7 amino acid residues have on LDM function and their possible contribution to the development of next-generation antifungals.
Methods: Three DNA cassettes generated using PCR with mutagenic oligonucleotide primers were transformed into yeast host strains. Liquid microdilution susceptibility assays were used to determine the minimal inhibitory drug concentrations of the mutant strains.
Results: None of the mutations were lethal and they did not appear to deleteriously affect enzyme function. Instead, these mutations increased susceptibility to short- and medium-tailed, but not long-tailed azole antifungals. Alanine mutagenesis of the other four conserved LBP residues of interest (Y72, F241, H381, and S382) was previously found to give similar phenotypes.
Conclusions: This project analysed amino acids conserved within the LBP of LDM of 10 important fungal pathogens. Mutational analysis has helped us understand the impact these 7 amino acid residues have on LDM function and their possible contribution to the development of next-generation antifungals.