Abstract
In recent years, an approach called "chemical genetics" has been adopted in drug research to discover and validate new targets and to identify and optimize leads by high throughput screening. In this work, we tested the ability of a library of small peptidomimetics to induce phenotypic effects with functional implications on a panel of strains of the budding yeast Saccharomyces cerevisiae, both wild type and mutants, for respiratory function and multidrug resistance. Further elucidation of the function of these peptidomimetics was assessed by testing the effects of the compound with the most prominent inhibitory activity, 089, on gene expression using DNA microarrays. Pathway analysis showed the involvement of such a molecule in inducing oxidative damage through alterations in mitochondrial functions. Transcriptional experiments were confirmed by increased levels of ROS and activation of mitochondrial membrane potential. Our results demonstrate the influence of a functional HAP1 gene in the performance of S. cerevisiae as a model system.
Highlights
We selected a panel of strains in which it is possible to investigate the influence of a library of molecules on growth rate, respiratory metabolism (HAP1), and multidrug-membrane function (ERG6, SNQ2, and PDR3)
Chemical Genetics of S. cerevisiae bicyclic scaffold based on the 6,8-dioxa-3-azabicyclo[3.2.1]octane core and diverse substituents, in order to hypothesize a connection between specific groups and biological effects
Selection of the Panel of Strains—Several detectable phenotypes in the yeast cell are related to growth rate, respiratory metabolism, and cell wall-multidrug resistance
Summary
Molecules—In this study, a preconstituted library, composed of bicyclic peptidomimetics, bicycles from tartaric acid and amino acids (BTAa), was used for assays on selected yeast strains (supplemental Table 1). Compounds responsible for the variation in cell growth during the exponential phase or in the OD650 value of the stationary phase or both were selected for further characterization These compounds were tested at concentrations of 1, 0.5, 0.4, 0.3, 0.2, and 0.1 mM. Strains were grown in YPD supplemented with the molecule at 0.3 mM concentration and monitored with the same protocol applied in the first level assay. Cells were grown in YPD supplemented with 089 at concentrations of 1, 0.5, 0.4, 0.2, and 0.1 mM; a positive control was carried out without the compound. Yeast cells (BY4742, BY4742⌬erg, and BY4742⌬snq strains) were treated in YPD (pH 4.8) supplemented with 089 at 0.2 mM concentration and incubated at 28 °C with shaking for 4 h. Each aliquot was treated with Calcofluor White (M2R) (blue) to show the cell wall in order to count total cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
