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J. Biol. Chem., Vol. 277, Issue 24, 21254-21260, June 14, 2002
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From the Departments of The Gal4p family of yeast zinc cluster proteins
comprises over 50 members that are putative transcriptional regulators.
For example, Pdr1p and Pdr3p activate multidrug resistance genes by binding to pleiotropic drug response elements (PDREs) found in promoters of target genes such as PDR5, encoding a drug
efflux pump involved in resistance to cycloheximide. However, the role of many zinc cluster proteins is unknown. We tested a panel of strains
carrying deletions of zinc cluster genes in the presence of various
drugs. One deletion strain (
New Regulators of Drug Sensitivity in the Family of Yeast Zinc
Cluster Proteins*
and§¶
Medicine,
§ Biochemistry, and ¶ Microbiology and Immunology,
McGill University Health Centre, Royal Victoria Hospital, McGill
University, Montréal, Québec H3A 1A1, Canada
rdr1) was resistant to cycloheximide, whereas eight strains showed sensitivity to the antifungal ketoconazole or cycloheximide. Unnamed zinc cluster genes
identified in our screen were called RDS for regulators of
drug sensitivity. RNA levels of multidrug resistance genes such as
PDR16, SNQ2, and PDR5 were
decreased in many deletion strains. For example, cycloheximide
sensitivity of a stb5 strain was correlated with
decreased RNA levels and promoter activity of the PDR5
gene. We tested if activation of PDR5 is mediated via a
PDRE by inserting this DNA element in front of a minimal promoter
linked to the lacZ gene. Strikingly, activity of the reporter was decreased in a stb5 strain. The purified
DNA binding domain of Stb5p bound to a PDRE in vitro.
Mutations in the PDRE known to affect binding of Pdr1p/Pdr3p showed
similar effects when assayed with Stb5p. These results strongly suggest
that Stb5p is a transcriptional activator of multidrug resistance
genes. Thus, we have identified new regulators of drug
sensitivity in the family of zinc cluster proteins.
*
This work was supported in part by grants from the Canadian
Institute of Health Research of Canada (Genomics) and the National Sciences and Engineering Research Council of Canada (to B. T.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Scholar of the Fonds de la Recherche en Santé du
Québec. To whom correspondence should be addressed: Dept. of
Medicine, McGill University Health Centre, Royal Victoria Hospital, 687 Pine Ave. West, Montréal, Québec H3A 1A1, Canada.
Tel.: 514-842-1231 (ext. 35046); Fax: 514-982-0893; E-mail:
turcotte@lan1.molonc.mcgill.ca.
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