Identification of Gold Nanoparticle-Resistant Mutants of Saccharomyces cerevisiae Suggests a Role for Respiratory Metabolism in Mediating Toxicity

Smith, Mark R.; Boenzli, Matthew G.; Hindagolla, Vihangi; Ding, Jun; Miller, John M.; Hutchison, James E.; Greenwood, Jeffrey A.; Abeliovich, Hagai; Bakalinsky, Alan T.

doi:10.1128/aem.01737-12

Cited by 19 publications

(18 citation statements)

References 31 publications

(33 reference statements)

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“…This analysis confirmed similar autofluorescence (Smith et al 2013) of the wild-type and PEP3 -overexpression strains, but indicated greater FM4-64 fluorescence intensity in the PEP3 -overexpression strain, 390 vs 108 arbitrary units (Fig. 3).…”

Section: Resultssupporting

confidence: 71%

PEP3 overexpression shortens lag phase but does not alter growth rate in Saccharomyces cerevisiae exposed to acetic acid stress

Ding

Holzwarth

Bradford

et al. 2015

Appl Microbiol Biotechnol

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In fungi, two recognized mechanisms contribute to pH homeostasis: the plasma membrane proton-pumping ATPase that exports excess protons and the vacuolar proton-pumping ATPase (V-ATPase) that mediates vacuolar proton uptake. Here, we report that overexpression of PEP3 which encodes a component of the HOPS and CORVET complexes involved in vacuolar biogenesis, shortened lag phase in Saccharomyces cerevisiae exposed to acetic acid stress. By confocal microscopy, PEP3-overexpressing cells stained with the vacuolar membrane-specific dye, FM4-64 had more fragmented vacuoles than the wild-type control. The stained overexpression mutant was also found to exhibit about 3.6-fold more FM4-64 fluorescence than the wild-type control as determined by flow cytometry. While the vacuolar pH of the wild-type strain grown in the presence of 80 mM acetic acid was significantly higher than in the absence of added acid, no significant difference was observed in vacuolar pH of the overexpression strain grown either in the presence or absence of 80 mM acetic acid. Based on an indirect growth assay, the PEP3-overexpression strain exhibited higher V-ATPase activity. We hypothesize that PEP3 overexpression provides protection from acid stress by increasing vacuolar surface area and V-ATPase activity and, hence, proton-sequestering capacity.

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Section: Resultssupporting

confidence: 71%

PEP3 overexpression shortens lag phase but does not alter growth rate in Saccharomyces cerevisiae exposed to acetic acid stress

Ding

Holzwarth

Bradford

et al. 2015

Appl Microbiol Biotechnol

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“…Recent reports on the reactivity of Au NPs focused on their effects on biocompatibility, uptake, toxicity, and mutagenic effects. [26][27][28][29][30] However, in general, there is a lack of conclusive information on the size as the key factor for Au NPs cytotoxicity and genotoxicity, and on the relationship between size, mass concentration and the absolute particle number.…”

Section: Discussionmentioning

confidence: 99%

Aneuploidogenic effects and DNA oxidation induced in vitro by differently sized gold nanoparticles

Bucchianico¹,

Fabbrizi²,

Cirillo³

et al. 2014

IJN

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Gold nanoparticles (Au NPs) are used in many fields, including biomedical applications; however, no conclusive information on their potential cytotoxicity and genotoxicity mechanisms is available. For this reason, experiments in human primary lymphocytes and murine macrophages (Raw264.7) were performed exposing cells to spherical citrate-capped Au NPs with two different nominal diameters (5 nm and 15 nm). The proliferative activity, mitotic, apoptotic, and necrotic markers, as well as chromosomal damage were assessed by the cytokinesis-block micronucleus cytome assay. Fluorescence in situ hybridization with human and murine pancentromeric probes was applied to distinguish between clastogenic and aneuploidogenic effects. Our results indicate that 5 nm and 15 nm Au NPs are able to inhibit cell proliferation by apoptosis and to induce chromosomal damage, in particular chromosome mis-segregation. DNA strand breaks were detected by comet assay, and the modified protocol using endonuclease-III and formamidopyrimidine-DNA glycosylase restriction enzymes showed that pyrimidines and purines were oxidatively damaged by Au NPs. Moreover, we show a size-independent correlation between the cytotoxicity of Au NPs and their tested mass concentration or absolute number, and genotoxic effects which were more severe for Au NP 15 nm compared to Au NP 5 nm. Results indicate that apoptosis, aneuploidy, and DNA oxidation play a pivotal role in the cytotoxicity and genotoxicity exerted by Au NPs in our cell models.

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“…When exposure occurred in YP‐galactose, Cu salt had significantly greater effect on metabolic activity (IC50: 0.96 mg Cu/L) compared with the 28 nm CuO NPs (IC50: 70.3 mg Cu/L) which were subsequently more toxic than 64 nm CuO NPs (IC50: 172.6 mg Cu/L). Saccharomyces cerevisiae under respiratory conditions were more sensitive to CuO NPs exposures compared with fermentative conditions which may implicate respiratory metabolism or the mitochondria in facilitating CuO NP toxicity, as suggested with gold NPs , however we have no direct evidence if this is the case. The lower toxic effect of CuO NPs compared with Cu salts has been shown in the gram‐negative bacteria Escherichia coli and Vibrio fischeri , the protozoa Tetrahymena thermophila , and the freshwater crustacean Thamnocephalus platyurus .…”

Section: Discussionmentioning

confidence: 79%

Copper oxide nanoparticles inhibit the metabolic activity of Saccharomyces cerevisiae

Mashock

Kappell

Hallaj

2015

Enviro Toxic and Chemistry

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Copper oxide nanoparticles (CuO NPs) are used increasingly in industrial applications and consumer products and thus may pose risk to human and environmental health. The interaction of CuO NPs with complex media and the impact on cell metabolism when exposed to sublethal concentrations are largely unknown. In the present study, the short-term effects of 2 different sized manufactured CuO NPs on metabolic activity of Saccharomyces cerevisiae were studied. The role of released Cu(2+) during dissolution of NPs in the growth media and the CuO nanostructure were considered. Characterization showed that the 28 nm and 64 nm CuO NPs used in the present study have different primary diameter, similar hydrodynamic diameter, and significantly different concentrations of dissolved Cu(2+) ions in the growth media released from the same initial NP mass. Exposures to CuO NPs or the released Cu(2+) fraction, at doses that do not have impact on cell viability, showed significant inhibition on S. cerevisiae cellular metabolic activity. A greater CuO NP effect on the metabolic activity of S. cerevisiae growth under respiring conditions was observed. Under the tested conditions the observed metabolic inhibition from the NPs was not explained fully by the released Cu ions from the dissolving NPs.

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Identification of Gold Nanoparticle-Resistant Mutants of Saccharomyces cerevisiae Suggests a Role for Respiratory Metabolism in Mediating Toxicity

Cited by 19 publications

References 31 publications

PEP3 overexpression shortens lag phase but does not alter growth rate in Saccharomyces cerevisiae exposed to acetic acid stress

PEP3 overexpression shortens lag phase but does not alter growth rate in Saccharomyces cerevisiae exposed to acetic acid stress

Aneuploidogenic effects and DNA oxidation induced in vitro by differently sized gold nanoparticles

Copper oxide nanoparticles inhibit the metabolic activity of Saccharomyces cerevisiae

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