Open AccessIsolation and functional analysis of the Kl PDR 16 gene
Author(s) -
Goffa Eduard,
Balazfyova Zuzana,
Toth Hervay Nora,
Simova Zuzana,
Balazova Maria,
Griac Peter,
Gbelska Yvetta
Publication year - 2014
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/1567-1364.12102
Subject(s) - biology , kluyveromyces lactis , oligomycin , efflux , rhodamine 123 , yeast , mutant , saccharomyces cerevisiae , biochemistry , gene , microbiology and biotechnology , ethidium bromide , rhodamine 6g , ergosterol , multiple drug resistance , drug resistance , atpase , dna , chemistry , enzyme , organic chemistry , molecule
The fight against multidrug‐resistant pathogens requires an understanding of the underlying cellular mechanisms. In this work, we isolate and characterize one of the multidrug resistance determinants in K luyveromyces lactis, the Kl PDR 16 gene. We show that Kl Pdr16p (345 aa), which belongs to the Kl Pdr1p regulon, is a functional homologue of the S accharomyces cerevisiae Pdr16p. Deletion of Kl PDR 16 resulted in hypersensitivity of K . lactis cells to antifungal azoles, oligomycin, rhodamine 6G, 4‐nitroquinoline‐N‐oxide and alkali metal cations. The Klpdr16∆ mutation led to a decreased content of ergosterol in whole‐cell extract. In spite of the hypersensitivity of Klpdr16∆ mutant cells to rhodamine 6G and oligomycin, the transcript level of the Kl PDR 5 gene and the rhodamine 6G efflux in the mutant was the same as in the parental strain. Increased accumulation of rhodamine 6G in Klpdr16∆ cells indicates that Kl PDR 16 limits the rate of passive drug diffusion across the membrane, without affecting the glucose‐induced drug export. The results obtained show that Kl PDR 16, similar to its orthologues in other yeast species, influences the passive drug diffusion into the yeast cell.