A Genome-Wide Screening in Saccharomyces cerevisiae for Genes That Confer Resistance to the Anticancer Agent Cisplatin

Burger, Herman; Capello, Astrid; Schenk, Paul W.; Stoter, G.; Brouwer, Jaap; Nooter, Kees

doi:10.1006/bbrc.2000.2361

Cited by 36 publications

(33 citation statements)

References 31 publications

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“…Sensitivity to cytotoxic chemicals was determined by a semiquantitative spot assay as described previously (Burger et al, 2000). Briefly, S. cerevisiae cells from freshly streaked plates were inoculated and grown to mid-log phase in selective liquid YNB medium.…”

Section: Methodsmentioning

confidence: 99%

“…Sensitivity to ionizing radiation was tested by a similar assay, during which spotting of the yeast cells was immediately followed by irradiation with increasing doses of ␥-rays using opposing 137 Cs sources (Gamma Cell 40; Atomic Energy of Canada, Ottawa, Canada) at a rate of 1.06 to 1.08 Gy/min. Sensitivity to ultraviolet light was also tested by a semiquantitative spot assay as described previously (Burger et al, 2000), using a germicidal lamp at 254 nm. Sensitivity to cisplatin and doxorubicin was analyzed in detail by a quantitative clonogenic survival assay.…”

Section: Methodsmentioning

confidence: 99%

“…Serial dilutions of mid-log phase yeast cells were plated onto selective YNB containing various drug concentrations. Plates were incubated at 30°C for 3 to 4 days, colonies were counted, and percentage survival was calculated based on the number of colonies arising in the absence of cytotoxic agents (Burger et al, 2000;Schenk et al, 2002). Where appropriate, the slopes of the log-linear concentration-survival curves were determined by linear regression analysis using SPSS 10.1 (SPSS Inc., Chicago, IL).…”

Section: Methodsmentioning

confidence: 99%

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Anticancer Drug Resistance Induced by Disruption of the Saccharomyces cerevisiae NPR2 Gene: a Novel Component Involved in Cisplatin- and Doxorubicin-Provoked Cell Kill

Schenk

Brok²,

Boersma³

et al. 2003

Mol Pharmacol

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The therapeutic potential of antitumor drugs is seriously limited by the manifestation of cellular drug resistance. We used the budding yeast Saccharomyces cerevisiae as a model system to identify novel mechanisms of resistance to one of the most active anticancer agents, cisplatin. We pinpointed NPR2 (nitrogen permease regulator 2) as a gene whose disruption conferred resistance to cisplatin. In addition, we observed a 4-fold cross-resistance of yeast npr2⌬ cells (i.e., cells from which the NPR2 gene had been disrupted) to the anticancer drug doxorubicin, in combination with hypersensitivity to cadmium chloride. Furthermore, npr2⌬ cells displayed unaltered cellular cisplatin and doxorubicin accumulation and showed an enhanced rate of spontaneous mutation compared with the isogenic parent. These data indicate that the npr2⌬ phenotype overlaps that of the sky1⌬ cells that we characterized previously (Mol Pharmacol 61: 659 -666, 2002). Therefore, we generated yeast npr2⌬ sky1⌬ double-knockout cells and performed clonogenic survival assays for cisplatin and doxorubicin, which revealed that NPR2 and SKY1 (SR-protein-specific kinase from budding yeast) are epistatic. The double-knockout strain was just as resistant to cisplatin and doxorubicin as the single-knockout strain that was most resistant to either drug. In conclusion, we identified NPR2 as a novel component involved in cell kill provoked by cisplatin and doxorubicin, and our data support the hypothesis that NPR2 and SKY1 may use mutual regulatory routes to mediate the cytotoxicity of these anticancer drugs.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Anticancer Drug Resistance Induced by Disruption of the Saccharomyces cerevisiae NPR2 Gene: a Novel Component Involved in Cisplatin- and Doxorubicin-Provoked Cell Kill

Schenk

Brok²,

Boersma³

et al. 2003

Mol Pharmacol

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“…The genes discovered in our study fall into several functional groups including nucleotide metabolism, protein metabolism, and regulation of cell proliferation. These functions have also been implicated by genetic screens for cisplatin resistance in yeast (24,29,41) and by transcriptional profiling of human cells (30)(31)(32). These correlated findings, made in organisms from three different taxa (7), suggest that some of the cisplatin response mechanisms are conserved between D. discoideum and humans.…”

Section: Discussionmentioning

confidence: 88%

Global transcriptional responses to cisplatin in Dictyostelium discoideum identify potential drug targets

Driessche¹,

Alexander

Kuspa

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Dictyostelium discoideum is a useful model for studying mechanisms of cisplatin drug sensitivity. Our previous findings, that mutations in sphingolipid metabolism genes confer cisplatin resistance in D. discoideum and in human cells, raised interest in the resistance mechanisms and their implications for cisplatin chemotherapy. Here we used expression microarrays to monitor physiological changes and to identify pathways that are affected by cisplatin treatment of D. discoideum. We found >400 genes whose regulation was altered by cisplatin treatment of wild-type cells, including groups of genes that participate in cell proliferation and in nucleotide and protein metabolism, showing that the cisplatin response is orderly and multifaceted. Transcriptional profiling of two isogenic cisplatin-resistant mutants, impaired in different sphingolipid metabolism steps, showed that the effect of cisplatin treatment was greater than the effect of the mutations, indicating that cisplatin resistance in the mutants is due to specific abilities to overcome the drug effects rather than to general drug insensitivity. Nevertheless, the mutants exhibited significantly different responses to cisplatin compared with the parent, and >200 genes accounted for that difference. Mutations in five cisplatin response genes (sgkB, csbA, acbA, smlA, and atg8) resulted in altered drug sensitivity, implicating novel pathways in cisplatin response. Our data illustrate how modeling complex cellular responses to drugs in genetically stable and tractable systems can uncover new targets with the potential for improving chemotherapy.drug resistance ͉ sphingolipids ͉ sphingosine kinase ͉ sphingosine-1-phosphate lyase ͉ transcriptional profiling D ictyostelium discoideum is well known for studies of cell and developmental biology and is gaining interest as a model for studying human disease, including cancer (1). A substantial body of work has accumulated using D. discoideum to identify genes and proteins associated with resistance to the anti-cancer drug cisplatin (2).Cisplatin [cis-diamminedichloroplatinum(II)] is used to treat a variety of solid tumors (3), but efficacy is often limited by drug resistance (4). Studies in human tumor cells and cell lines have implicated a number of resistance mechanisms and demonstrated that resistance is multifaceted (5).D. discoideum provides a genetically tractable system to the problem of cisplatin resistance. It is a eukaryote with cellular properties similar to those of human cells (6). Its genome is sequenced, and many genes are homologous to human genes (7). Using insertional mutagenesis screens (8), we isolated mutants with single gene disruptions that have decreased sensitivity to cisplatin (9). One of these strains was null for the sphingosine-1-phosphate (S-1-P) lyase gene sglA, establishing the first link between sphingolipid metabolism and cisplatin sensitivity and defining S-1-P lyase as a potential target for controlling cisplatin sensitivity.S-1-P synthesis is catalyzed by sphingosine kinase from sphin...

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“…Thus, ZDS1 was isolated as a multicopy suppressor of sensitivity to the calmodulin inhibitor TFP in the ssd1D strain (Tsuchiya et al 1996) as well as a multicopy suppressor of the hsl1 and hsl7 mutants (Ma et al 1996), of cdc28-1N (Yuet al 1996), and of a tif1 mutant (tif1 encodes the translation initiation factor eIF-4A) (Schwer et al 1998). ZDS2 was isolated as a multicopy suppressor of sin4, a gene that confers resistance to the anticancer agent cisplatin (Burger et al 2000). ZDS1 and ZDS2 were also isolated as (i) multicopy suppressors of temperature-sensitive mutations of a yeast mRNA-capping enzyme (CEG1) (Schwer and Shuman 1996), (ii) negative regulators of Cdc42p (Bi and Pringle 1996), (iii) multicopy suppressors of a cka2 mutant (cka2 encodes the a9-subunit of casein kinase II) (Bandhakavi et al 2003), (iv) suppressors of the camptothecin-hypersensitive trf4 mutant (Walowsky et al 1999), (v) genes that stabilize short linear centromeric plasmids (Roy and Runge 1999), (vi) multicopy suppressors of an rhc21 mutant (rhc21 encodes a component of the cohesin complex) (Heo et al 1999), (vii) genes that interact with the Bcy1p N-terminal domain (Griffioen et al 2001), and (viii) multicopy suppressors of a 1, 3-b-glucan synthase mutant (Sekiya-Kawasaki et al 2002).…”

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confidence: 99%

zds1, a Novel Gene Encoding an Ortholog of Zds1 and Zds2, Controls Sexual Differentiation, Cell Wall Integrity and Cell Morphology in Fission Yeast

Yakura¹,

Ozoe²,

Ishida³

et al. 2006

Genetics

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While screening for genes that reverse the sporulation-deficient phenotype of the ras1D diploid Schizosaccharomyces pombe strain, we identified zds1. This gene shares sequence homology with the ZDS1 and ZDS2 genes from Saccharomyces cerevisiae, which appear to be involved in multiple cellular events. Expression of Zds1 in ras1D diploid cells elevated their sporulation rate from 0.3 to 11.2%. Expression of the Zds1 C-terminal region increased the sporulation rate further (to 21.9%) while introduction of the Zds1 N-terminal region had no effect. zds1 expression did not induce sporulation in strains with mutations in genes participating in the downstream MAP kinase cascade. The zds1-disrupted strain is sensitive to CaCl 2 , and this effect is suppressed by the C-terminal region of Zds1. The growth of the zds1D strain is markedly inhibited by cold temperatures, while its viability decreased in the stationary phase. Moreover, the zds1D strain is round in shape and very sensitive to zymolyase, and its cell wall becomes thicker than that of wild type. Thus, zds1 must be required to maintain cell wall integrity. The Zds1-GFP fusion protein localized to the cytosol, the septum, and the cell cortex. Its localization in the septum was dependent on its C-terminal region. Overexpression of the C-terminal region of Zds1 induced multi-septa and abnormal zygotes. We propose that the C-terminal region is the functional domain of Zds1 while the N-terminal region is a negative regulatory region. Thus, Zds1 is involved in multiple cellular events in fission yeast, including sexual differentiation, Ca 21 tolerance, cell wall integrity, viability in the stationary phase, and cell morphology.

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A Genome-Wide Screening in Saccharomyces cerevisiae for Genes That Confer Resistance to the Anticancer Agent Cisplatin

Cited by 36 publications

References 31 publications

Anticancer Drug Resistance Induced by Disruption of the Saccharomyces cerevisiae NPR2 Gene: a Novel Component Involved in Cisplatin- and Doxorubicin-Provoked Cell Kill

Anticancer Drug Resistance Induced by Disruption of the Saccharomyces cerevisiae NPR2 Gene: a Novel Component Involved in Cisplatin- and Doxorubicin-Provoked Cell Kill

Global transcriptional responses to cisplatin in Dictyostelium discoideum identify potential drug targets

zds1, a Novel Gene Encoding an Ortholog of Zds1 and Zds2, Controls Sexual Differentiation, Cell Wall Integrity and Cell Morphology in Fission Yeast

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