Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput

• Autor(a) principal: Almeida, Ludimila Dias
• Data de Publicação: 2021
• Outros Autores: Silva, Ali Salim Faraj, Mota, Daniel Calixto, Vasconcelos, Adrielle Ayumi, Camargo, Antônio Pedro, Pires, Gabriel Silva, Furlan, Monique, Da Cunha Freire, Helena Martins Ribeiro, Klippel, Angélica Hollunder [UNESP], Silva, Suélen Fernandes [UNESP], Zanelli, Cleslei Fernando [UNESP], Carazzolle, Marcelo Falsarella, Oliver, Stephen G., Bilsland, Elizabeth
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://dx.doi.org/10.1128/mbio.03221-21; http://hdl.handle.net/11449/231580
• topic: Drug efflux; Drug transport; Drug uptake; Genetic interactions; Nonessential transporter double-deletion library; Plasma membrane transporter; Saccharomyces cerevisiae; Xenobiotics; Yeast

The 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.