A chemical genetic screen for modulators of asymmetrical 2,2'-dimeric naphthoquinones cytotoxicity in yeast

Autor: Yolanda M. Fortenberry, Milena Vuica-Ross, Kathleen Cowan, Ashkan Emadi, Ashley E. Ross
Jazyk: angličtina
Rok vydání: 2010
Předmět:
Saccharomyces cerevisiae Proteins
Cell Biology/Microbial Physiology and Metabolism
Mutant
Saccharomyces cerevisiae
Genetics and Genomics/Pharmacogenomics
lcsh:Medicine
Mitochondrion
Biology
Isomerism
Genetic Testing
Genetics and Genomics/Genomics
Inner mitochondrial membrane
lcsh:Science
Cell Biology/Chemical Biology of the Cell
Membrane Potential
Mitochondrial
Genetics
Microbial Viability
Multidisciplinary
Genetics and Genomics/Functional Genomics
lcsh:R
Wild type
Reproducibility of Results
Cell Biology/Cellular Death and Stress Responses
biology.organism_classification
Aerobiosis
Yeast
Culture Media
Mitochondria
Biochemistry
Fermentation
lcsh:Q
NAD+ kinase
Genome
Fungal
Genetics and Genomics/Comparative Genomics
Reactive Oxygen Species
Naphthoquinones
Research Article
Pharmacology/Drug Development
Genetic screen
Zdroj: PLoS ONE, Vol 5, Iss 5, p e10846 (2010)
PLoS ONE
ISSN: 1932-6203
Popis: BACKGROUND: Dimeric naphthoquinones (BiQ) were originally synthesized as a new class of HIV integrase inhibitors but have shown integrase-independent cytotoxicity in acute lymphoblastic leukemia cell lines suggesting their use as potential anti-neoplastic agents. The mechanism of this cytotoxicity is unknown. In order to gain insight into the mode of action of binaphthoquinones we performed a systematic high-throughput screen in a yeast isogenic deletion mutant array for enhanced or suppressed growth in the presence of binaphthoquinones. METHODOLOGY/PRINCIPAL FINDINGS: Exposure of wild type yeast strains to various BiQs demonstrated inhibition of yeast growth with IC(50)s in the microM range. Drug sensitivity and resistance screens were performed by exposing arrays of a haploid yeast deletion mutant library to BiQs at concentrations near their IC(50). Sensitivity screens identified yeast with deletions affecting mitochondrial function and cellular respiration as having increased sensitivity to BiQs. Corresponding to this, wild type yeast grown in the absence of a fermentable carbon source were particularly sensitive to BiQs, and treatment with BiQs was shown to disrupt the mitochondrial membrane potential and lead to the generation of reactive oxygen species (ROS). Furthermore, baseline ROS production in BiQ sensitive mutant strains was increased compared to wild type and could be further augmented by the presence of BiQ. Screens for resistance to BiQ action identified the mitochondrial external NAD(P)H dehydrogenase, NDE1, as critical to BiQ toxicity and over-expression of this gene resulted in increased ROS production and increased sensitivity of wild type yeast to BiQ. CONCLUSIONS/SIGNIFICANCE: In yeast, binaphthoquinone cytotoxicity is likely mediated through NAD(P)H:quonine oxidoreductases leading to ROS production and dysfunctional mitochondria. Further studies are required to validate this mechanism in mammalian cells.
Databáze: OpenAIRE
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