Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput.
View / Open Files
Authors
Almeida, Ludimila Dias
Silva, Ali Salim Faraj
Mota, Daniel Calixto
Vasconcelos, Adrielle Ayumi
Camargo, Antônio Pedro
Pires, Gabriel Silva
Furlan, Monique
Freire, Helena Martins Ribeiro da Cunha
Klippel, Angélica Hollunder
Silva, Suélen Fernandes
Zanelli, Cleslei Fernando
Carazzolle, Marcelo Falsarella
Publication Date
2021-12-21Journal Title
mBio
ISSN
2161-2129
Publisher
American Society for Microbiology
Volume
12
Issue
6
Language
eng
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Almeida, L. D., Silva, A. S. F., Mota, D. C., Vasconcelos, A. A., Camargo, A. P., Pires, G. S., Furlan, M., et al. (2021). Yeast Double Transporter Gene Deletion Library for Identification of Xenobiotic Carriers in Low or High Throughput.. mBio, 12 (6) https://doi.org/10.1128/mbio.03221-21
Abstract
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. IMPORTANCE Our library of double transporter deletion strains is a powerful tool for rapid identification of potential drug import and export routes, which can aid in determining the chemical groups necessary for transport via specific carriers. This information may be translated into a better design of drugs for optimal absorption by target tissues and the development of drugs whose utility is less likely to be compromised by the selection of resistant mutants.
Keywords
Saccharomyces cerevisiae, Yeast, Xenobiotics, Drug Transport, Drug Uptake, Genetic Interactions, Drug Efflux, Plasma Membrane Transporter, Nonessential Transporter Double-deletion Library
Sponsorship
Bill & Melinda Gates Foundation
FAPESP
Funder references
Biotechnology and Biological Sciences Research Council (BB/F008228/1)
Bill & Melinda Gates Foundation (OPP1087646)
Identifiers
34903049, PMC8669479
External DOI: https://doi.org/10.1128/mbio.03221-21
This record's URL: https://www.repository.cam.ac.uk/handle/1810/333250
Statistics
Total file downloads (since January 2020). For more information on metrics see the
IRUS guide.
Recommended or similar items
The current recommendation prototype on the Apollo Repository will be turned off on 03 February 2023. Although the pilot has been fruitful for both parties, the service provider IKVA is focusing on horizon scanning products and so the recommender service can no longer be supported. We recognise the importance of recommender services in supporting research discovery and are evaluating offerings from other service providers. If you would like to offer feedback on this decision please contact us on: support@repository.cam.ac.uk