酵母突變體itr1Δ (BY4741; YDR497c::kanMX4)

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.

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