Almeida, Ludimila DiasSilva, Ali Salim FarajMota, Daniel CalixtoVasconcelos, Adrielle AyumiCamargo, Antônio PedroPires, Gabriel SilvaFurlan, MoniqueDa Cunha Freire, Helena Martins RibeiroKlippel, Angélica Hollunder [UNESP]Silva, Suélen Fernandes [UNESP]Zanelli, Cleslei Fernando [UNESP]Carazzolle, Marcelo FalsarellaOliver, Stephen G.Bilsland, Elizabeth2022-04-292022-04-292021-12-01mBio, v. 12, n. 6, 2021.2150-75112161-2129http://hdl.handle.net/11449/231580The routes of uptake and efflux should be considered when developing new drugs so that they can effectively address their intracellular targets. As a general rule, drugs appear to enter cells via protein carriers that normally carry nutrients or metabolites. A previously developed pipeline that searched for drug transporters using Saccharomyces cerevisiae mutants carrying single-gene deletions identified import routes for most compounds tested. However, due to the redundancy of transporter functions, we propose that this methodology can be improved by utilizing double mutant strains in both low- and high-throughput screens. We constructed a library of over 14,000 strains harboring double deletions of genes encoding 122 nonessential plasma membrane transporters and performed low- and high-throughput screens identifying possible drug import routes for 23 compounds. In addition, the high-throughput assay enabled the identification of putative efflux routes for 21 compounds. Focusing on azole antifungals, we were able to identify the involvement of the myo-inositol transporter, Itr1p, in the uptake of these molecules and to confirm the role of Pdr5p in their export.engDrug effluxDrug transportDrug uptakeGenetic interactionsNonessential transporter double-deletion libraryPlasma membrane transporterSaccharomyces cerevisiaeXenobioticsYeastArtigo10.1128/mbio.03221-212-s2.0-85121978014