A biosensor-based approach reveals links between efflux pump expression and cell cycle regulation in pleiotropic drug resistance of yeast

Jian Li, Kristen Kolberg, Ulrich Schlecht, Robert P. Onge, Ana Aparicio, Joe Horecka, Ronald W. Davis, Maureen E. Hillenmeyer, Colin J.B. Harvey

Research output: Contribution to journalArticle

Abstract

Multidrug resistance is highly conserved in mammalian, fungal, and bacterial cells, is characterized by resistance to several unrelated xenobiotics, and poses significant challenges to managing infections and many cancers. Eukaryotes use a highly conserved set of drug efflux transporters that confer pleiotropic drug resistance (PDR). To interrogate the regulation of this critical process, here we developed a small molecule-responsive biosensor that couples transcriptional induction of PDR genes to growth rate in the yeast Saccharomyces cerevisiae. Using diverse PDR inducers and the homozygous diploid deletion collection, we applied this biosensor system to genome-wide screens for potential PDR regulators. In addition to recapitulating the activity of previously known factors, these screens identified a series of genes involved in a variety of cellular processes with significant but previously uncharacterized roles in the modulation of yeast PDR. Genes identified as down-regulators of the PDR included those encoding the MAD family of proteins involved in the mitotic spindle assembly checkpoint (SAC) complex. Of note, we demonstrated that genetic disruptions of the mitotic spindle assembly checkpoint elevate expression of PDR-mediating efflux pumps in response to exposure to a variety of compounds that themselves have no known influence on the cell cycle. These results not only establish our biosensor system as a viable tool for investigating PDR in a high-throughput fashion, but also uncover critical control mechanisms governing the PDR response and a previously uncharacterized link between PDR and cell cycle regulation in yeast.

Original languageEnglish (US)
Pages (from-to)1257-1266
Number of pages10
JournalJournal of Biological Chemistry
Volume294
Issue number4
DOIs
StatePublished - Jan 1 2019

Fingerprint

Biosensing Techniques
Drug Resistance
Biosensors
Yeast
Cell Cycle
Yeasts
Cells
Pumps
M Phase Cell Cycle Checkpoints
Pharmaceutical Preparations
Genes
Multiple Drug Resistance
Xenobiotics
Eukaryota
Diploidy
Saccharomyces cerevisiae
Genome
Throughput
Modulation
Molecules

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A biosensor-based approach reveals links between efflux pump expression and cell cycle regulation in pleiotropic drug resistance of yeast. / Li, Jian; Kolberg, Kristen; Schlecht, Ulrich; Onge, Robert P.; Aparicio, Ana; Horecka, Joe; Davis, Ronald W.; Hillenmeyer, Maureen E.; Harvey, Colin J.B.

In: Journal of Biological Chemistry, Vol. 294, No. 4, 01.01.2019, p. 1257-1266.

Research output: Contribution to journalArticle

Li, Jian ; Kolberg, Kristen ; Schlecht, Ulrich ; Onge, Robert P. ; Aparicio, Ana ; Horecka, Joe ; Davis, Ronald W. ; Hillenmeyer, Maureen E. ; Harvey, Colin J.B. / A biosensor-based approach reveals links between efflux pump expression and cell cycle regulation in pleiotropic drug resistance of yeast. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 4. pp. 1257-1266.
@article{960d77a619c44127aec0fc65e3323981,
title = "A biosensor-based approach reveals links between efflux pump expression and cell cycle regulation in pleiotropic drug resistance of yeast",
abstract = "Multidrug resistance is highly conserved in mammalian, fungal, and bacterial cells, is characterized by resistance to several unrelated xenobiotics, and poses significant challenges to managing infections and many cancers. Eukaryotes use a highly conserved set of drug efflux transporters that confer pleiotropic drug resistance (PDR). To interrogate the regulation of this critical process, here we developed a small molecule-responsive biosensor that couples transcriptional induction of PDR genes to growth rate in the yeast Saccharomyces cerevisiae. Using diverse PDR inducers and the homozygous diploid deletion collection, we applied this biosensor system to genome-wide screens for potential PDR regulators. In addition to recapitulating the activity of previously known factors, these screens identified a series of genes involved in a variety of cellular processes with significant but previously uncharacterized roles in the modulation of yeast PDR. Genes identified as down-regulators of the PDR included those encoding the MAD family of proteins involved in the mitotic spindle assembly checkpoint (SAC) complex. Of note, we demonstrated that genetic disruptions of the mitotic spindle assembly checkpoint elevate expression of PDR-mediating efflux pumps in response to exposure to a variety of compounds that themselves have no known influence on the cell cycle. These results not only establish our biosensor system as a viable tool for investigating PDR in a high-throughput fashion, but also uncover critical control mechanisms governing the PDR response and a previously uncharacterized link between PDR and cell cycle regulation in yeast.",
author = "Jian Li and Kristen Kolberg and Ulrich Schlecht and Onge, {Robert P.} and Ana Aparicio and Joe Horecka and Davis, {Ronald W.} and Hillenmeyer, {Maureen E.} and Harvey, {Colin J.B.}",
year = "2019",
month = "1",
day = "1",
doi = "10.1074/jbc.RA118.003904",
language = "English (US)",
volume = "294",
pages = "1257--1266",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "4",

}

TY - JOUR

T1 - A biosensor-based approach reveals links between efflux pump expression and cell cycle regulation in pleiotropic drug resistance of yeast

AU - Li, Jian

AU - Kolberg, Kristen

AU - Schlecht, Ulrich

AU - Onge, Robert P.

AU - Aparicio, Ana

AU - Horecka, Joe

AU - Davis, Ronald W.

AU - Hillenmeyer, Maureen E.

AU - Harvey, Colin J.B.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Multidrug resistance is highly conserved in mammalian, fungal, and bacterial cells, is characterized by resistance to several unrelated xenobiotics, and poses significant challenges to managing infections and many cancers. Eukaryotes use a highly conserved set of drug efflux transporters that confer pleiotropic drug resistance (PDR). To interrogate the regulation of this critical process, here we developed a small molecule-responsive biosensor that couples transcriptional induction of PDR genes to growth rate in the yeast Saccharomyces cerevisiae. Using diverse PDR inducers and the homozygous diploid deletion collection, we applied this biosensor system to genome-wide screens for potential PDR regulators. In addition to recapitulating the activity of previously known factors, these screens identified a series of genes involved in a variety of cellular processes with significant but previously uncharacterized roles in the modulation of yeast PDR. Genes identified as down-regulators of the PDR included those encoding the MAD family of proteins involved in the mitotic spindle assembly checkpoint (SAC) complex. Of note, we demonstrated that genetic disruptions of the mitotic spindle assembly checkpoint elevate expression of PDR-mediating efflux pumps in response to exposure to a variety of compounds that themselves have no known influence on the cell cycle. These results not only establish our biosensor system as a viable tool for investigating PDR in a high-throughput fashion, but also uncover critical control mechanisms governing the PDR response and a previously uncharacterized link between PDR and cell cycle regulation in yeast.

AB - Multidrug resistance is highly conserved in mammalian, fungal, and bacterial cells, is characterized by resistance to several unrelated xenobiotics, and poses significant challenges to managing infections and many cancers. Eukaryotes use a highly conserved set of drug efflux transporters that confer pleiotropic drug resistance (PDR). To interrogate the regulation of this critical process, here we developed a small molecule-responsive biosensor that couples transcriptional induction of PDR genes to growth rate in the yeast Saccharomyces cerevisiae. Using diverse PDR inducers and the homozygous diploid deletion collection, we applied this biosensor system to genome-wide screens for potential PDR regulators. In addition to recapitulating the activity of previously known factors, these screens identified a series of genes involved in a variety of cellular processes with significant but previously uncharacterized roles in the modulation of yeast PDR. Genes identified as down-regulators of the PDR included those encoding the MAD family of proteins involved in the mitotic spindle assembly checkpoint (SAC) complex. Of note, we demonstrated that genetic disruptions of the mitotic spindle assembly checkpoint elevate expression of PDR-mediating efflux pumps in response to exposure to a variety of compounds that themselves have no known influence on the cell cycle. These results not only establish our biosensor system as a viable tool for investigating PDR in a high-throughput fashion, but also uncover critical control mechanisms governing the PDR response and a previously uncharacterized link between PDR and cell cycle regulation in yeast.

UR - http://www.scopus.com/inward/record.url?scp=85060661583&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85060661583&partnerID=8YFLogxK

U2 - 10.1074/jbc.RA118.003904

DO - 10.1074/jbc.RA118.003904

M3 - Article

VL - 294

SP - 1257

EP - 1266

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 4

ER -