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@ARTICLE{Tanabe:856857,
author = {Tanabe, Koichi and Bonus, Michele and Tomiyama, Susumu and
Miyoshi, Kunji and Nagi, Minoru and Niimi, Kyoko and
Chindamporn, Ariya and Gohlke, Holger and Schmitt, Lutz and
Cannon, Richard D. and Niimi, Masakazu and Lamping, Erwin},
title = {{FK}506 resistance of {S}accharomyces cerevisiae {P}dr5 and
{C}andida albicans {C}dr1 involves mutations in the
transmembrane domains and extracellular loops},
journal = {Antimicrobial agents and chemotherapy},
volume = {63},
number = {1},
issn = {1098-6596},
address = {Washington, DC},
publisher = {Soc.},
reportid = {FZJ-2018-06196},
pages = {e01146-18},
year = {2019},
abstract = {The 23-membered-ring macrolide tacrolimus, a commonly used
immunosuppressant, also known as FK506, is a broad-spectrum
inhibitor and an efflux pump substrate of pleiotropic drug
resistance (PDR) ATP-binding cassette (ABC) transporters.
Little, however, is known about the molecular mechanism by
which FK506 inhibits PDR transporter drug efflux. Thus, to
obtain further insights we searched for FK506-resistant
mutants of Saccharomyces cerevisiae cells overexpressing
either the endogenous multidrug efflux pump, Pdr5, or its
Candida albicans orthologue, Cdr1. A simple, but powerful,
screen gave 69 FK506-resistant mutants with, between them,
72 mutations in either Pdr5 (37) or Cdr1 (35). Twenty
mutations were in just three Pdr5/Cdr1 equivalent amino acid
positions T550/T540 and T552/S542 of extracellular loop 1
(EL1) and A723/A713 of EL3. Sixty of the 72 mutations were
either in the ELs or the extracellular halves of individual
transmembrane spans (TMSs), while 11 mutations were found
near the centre of individual TMSs, mostly in predicted
TMS-TMS contact points, and only two mutations were in the
cytosolic nucleotide-binding domains of Pdr5. We propose
that FK506 inhibits Pdr5 and Cdr1 drug efflux by slowing
transporter opening and/or substrate release, and that
FK506-resistance of Pdr5/Cdr1 drug efflux is achieved by
modifying critical intramolecular contact points that, when
mutated, enable the co-transport of FK506 with other pump
substrates. This may also explain why the 35 Cdr1 mutations
that caused FK506-insensitivity of fluconazole efflux
differed from the 13 Cdr1 mutations that caused
FK506-insensitivity of cycloheximide efflux.},
cin = {JSC / ICS-6 / NIC},
ddc = {610},
cid = {I:(DE-Juel1)JSC-20090406 / I:(DE-Juel1)ICS-6-20110106 /
I:(DE-Juel1)NIC-20090406},
pnm = {511 - Computational Science and Mathematical Methods
(POF3-511) / Forschergruppe Gohlke $(hkf7_20170501)$},
pid = {G:(DE-HGF)POF3-511 / $G:(DE-Juel1)hkf7_20170501$},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:30348662},
UT = {WOS:000454140200008},
doi = {10.1128/AAC.01146-18},
url = {https://juser.fz-juelich.de/record/856857},
}