High-Copy Overexpression Screening Reveals PDR5 as the Main Doxorubicin Resistance Gene in Yeast

Demir, Ayşe Banu; Koç, Ahmet

doi:10.1371/journal.pone.0145108

Cited by 10 publications

(7 citation statements)

References 54 publications

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“…Consistent with this conclusion, Sla1- and End3-defective mutants also exhibited elevated resistance to doxorubicin ( 43 ). Conversely, and also consistent with a role for Akl1 in preventing doxorubicin entry by blocking endocytosis, akl1 Δ cells are hypersensitive to this antibiotic ( 43 , 62 , 63 ).…”

Section: Resultssupporting

confidence: 59%

TOR Complex 2-Regulated Protein Kinase Fpk1 Stimulates Endocytosis via Inhibition of Ark1/Prk1-Related Protein Kinase Akl1 in Saccharomyces cerevisiae

Roelants

Leskoske

Pedersen

et al. 2017

Molecular and Cellular Biology

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Depending on the stress, plasma membrane alterations activate or inhibit yeast target of rapamycin (TOR) complex 2, which, in turn, upregulates or downregulates the activity of its essential downstream effector, protein kinase Ypk1. Through phosphorylation of multiple substrates, Ypk1 controls many processes that restore homeostasis. One such substrate is protein kinase Fpk1, which is negatively regulated by Ypk1. Fpk1 phosphorylates and stimulates flippases that translocate aminoglycerophospholipids from the outer to the inner leaflet of the plasma membrane. Fpk1 has additional roles, but other substrates were uncharacterized. We show that Fpk1 phosphorylates and inhibits protein kinase Akl1, related to protein kinases Ark1 and Prk1, which modulate the dynamics of actin patch-mediated endocytosis. Akl1 has two Fpk1 phosphorylation sites (Ark1 and Prk1 have none) and is hypophosphorylated when Fpk1 is absent. Conversely, under conditions that inactivate TORC2-Ypk1 signaling, which alleviates Fpk1 inhibition, Akl1 is hyperphosphorylated. Monitoring phosphorylation of known Akl1 substrates (Sla1 and Ent2) confirmed that Akl1 is hyperactive when not phosphorylated by Fpk1. Fpk1-mediated negative regulation of Akl1 enhances endocytosis, because an Akl1 mutant immune to Fpk1 phosphorylation causes faster dissociation of Sla1 from actin patches, confers elevated resistance to doxorubicin (a toxic compound whose entry requires endocytosis), and impedes Lucifer yellow uptake (a marker of fluid phase endocytosis). Thus, TORC2-Ypk1, by regulating Fpk1-mediated phosphorylation of Akl1, adjusts the rate of endocytosis.

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Section: Resultssupporting

confidence: 59%

TOR Complex 2-Regulated Protein Kinase Fpk1 Stimulates Endocytosis via Inhibition of Ark1/Prk1-Related Protein Kinase Akl1 in Saccharomyces cerevisiae

Roelants

Leskoske

Pedersen

et al. 2017

Molecular and Cellular Biology

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“…Meiosis and sporulation were observed to be significantly induced in response of yeast cells exposed to doxorubicin. Meiosis was shown to be up-regulated in the presence of doxorubicin in yeast cells 49 . A deficiency in glucose and nitrogen are known to induce meiosis followed G1 arrest of the mitotic cell cycle in diploid yeast cells 54 , 55 .…”

Section: Discussionmentioning

confidence: 99%

“…Detailed investigation of the transcriptional response indicated that the genes involved in cellular response to DNA damage and DNA repair were also induced. Several genes functioning in DNA repair were observed to be up-regulated upon doxorubicin exposure 49 . Modular analysis of DEGN also indicated that a module is significantly associated with nucleosome assembly/disassembly and several genes within this module was also found to be involved in DNA repair.…”

Section: Discussionmentioning

confidence: 99%

Doxorubicin induces an extensive transcriptional and metabolic rewiring in yeast cells

Taymaz-Nikerel

Karabekmez

Eraslan

et al. 2018

Sci Rep

134

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Doxorubicin is one of the most effective chemotherapy drugs used against solid tumors in the treatment of several cancer types. Two different mechanisms, (i) intercalation of doxorubicin into DNA and inhibition of topoisomerase II leading to changes in chromatin structure, (ii) generation of free radicals and oxidative damage to biomolecules, have been proposed to explain the mode of action of this drug in cancer cells. A genome-wide integrative systems biology approach used in the present study to investigate the long-term effect of doxorubicin in Saccharomyces cerevisiae cells indicated the up-regulation of genes involved in response to oxidative stress as well as in Rad53 checkpoint sensing and signaling pathway. Modular analysis of the active sub-network has also revealed the induction of the genes significantly associated with nucleosome assembly/disassembly and DNA repair in response to doxorubicin. Furthermore, an extensive re-wiring of the metabolism was observed. In addition to glycolysis, and sulfate assimilation, several pathways related to ribosome biogenesis/translation, amino acid biosynthesis, nucleotide biosynthesis, de novo IMP biosynthesis and one-carbon metabolism were significantly repressed. Pentose phosphate pathway, MAPK signaling pathway biological processes associated with meiosis and sporulation were found to be induced in response to long-term exposure to doxorubicin in yeast cells.

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“…Observations from other studies illustrate the higher resistance of S. cerevisiae against doxorubicin and the protective effect of glutathione and or metallothioneins [ 70 ]. The assessment of the results should be considered the phase of the cell cycle, which has a direct effect on the toxicity of doxorubicin [ 71 , 72 , 73 , 74 ]. Further investigations are necessary as well as a selective effect on healthy and prostate tumor tissue.…”

Section: Discussionmentioning

confidence: 99%

Zinc-Modified Nanotransporter of Doxorubicin for Targeted Prostate Cancer Delivery

et al. 2017

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This work investigated the preparation of chitosan nanoparticles used as carriers for doxorubicin for targeted cancer delivery. Prepared nanocarriers were stabilized and functionalized via zinc ions incorporated into the chitosan nanoparticle backbone. We took the advantage of high expression of sarcosine in the prostate cancer cells. The prostate cancer targeting was mediated by the AntiSar antibodies decorated surface of the nanocage. Formation of the chitosan nanoparticles was determined using a ninhydrin assay and differential pulse voltammetry. Obtained results showed the strong effect of tripolyphosphine on the nanoparticle formation. The zinc ions affected strong chitosan backbone coiling both in inner and outer chitosan nanoparticle structure. Zinc electrochemical signal depended on the level of the complex formation and the potential shift from −960 to −950 mV. Formed complex is suitable for doxorubicin delivery. It was observed the 20% entrapment efficiency of doxorubicin and strong dependence of drug release after 120 min in the blood environment. The functionality of the designed nanotransporter was proven. The purposed determination showed linear dependence in the concentration range of Anti-sarcosine IgG labeled gold nanoparticles from 0 to 1000 µg/mL and the regression equation was found to be y = 3.8x − 66.7 and R2 = 0.99. Performed ELISA confirmed the ability of Anti-sarcosine IgG labeled chitosan nanoparticles with loaded doxorubicin to bind to the sarcosine molecule. Observed hemolytic activity of the nanotransporter was 40%. Inhibition activity of our proposed nanotransporter was evaluated to be 0% on the experimental model of S. cerevisiae. Anti-sarcosine IgG labeled chitosan nanoparticles, with loaded doxorubicin stabilized by Zn ions, are a perspective type of nanocarrier for targeted drug therapy managed by specific interaction with sarcosine and metallothionein for prostate cancer.

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High-Copy Overexpression Screening Reveals PDR5 as the Main Doxorubicin Resistance Gene in Yeast

Cited by 10 publications

References 54 publications

TOR Complex 2-Regulated Protein Kinase Fpk1 Stimulates Endocytosis via Inhibition of Ark1/Prk1-Related Protein Kinase Akl1 in Saccharomyces cerevisiae

TOR Complex 2-Regulated Protein Kinase Fpk1 Stimulates Endocytosis via Inhibition of Ark1/Prk1-Related Protein Kinase Akl1 in Saccharomyces cerevisiae

Doxorubicin induces an extensive transcriptional and metabolic rewiring in yeast cells

Zinc-Modified Nanotransporter of Doxorubicin for Targeted Prostate Cancer Delivery

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