Assessing Yeast Cell Survival Following Hydrogen Peroxide Exposure

Tran, Katelynn; Green, Erin M.

doi:10.21769/bioprotoc.3149

Cited by 25 publications

(23 citation statements)

References 10 publications

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“…We first tested our growth conditions for wildtype cells grown in aerobic and hypoxic conditions. We observed that we obtained the most consistent results by diluting stationary phase cultures to very low OD 600 and allowing them to grow to OD 600 ∼0.4-0.8 over the course of 18 hours in hypoxia, similar to how we tested set4Δ mutants sensitivity to oxidative stress (10,12). Under these conditions, the PAU genes were highly upregulated, and there was also significant upregulation of YGL262W , although expression of COS12 and YPS5 at TEL07L remained mostly unchanged in hypoxic compared to aerobic growth ( Figure 2A ).…”

Section: Resultssupporting

confidence: 65%

“…For hypoxic growth, the culture flasks were placed in BD GasPak EZ anaerobe pouch system and incubated at 30°C. For hydrogen peroxide-treated cultures, cells were grown to mid-log phase (OD 600 ∼0.6-0.8) and then treated with 0.4mM H 2 O 2 for 30 min (12).…”

Section: Methodsmentioning

confidence: 99%

“…Our previous work identified a role for the yeast protein Set4, a paralog to Set3, in protecting cells during oxidative stress, primarily through gene expression regulation of stress response genes (10). Set4 is lowly expressed in yeast cells under standard laboratory growth conditions, although deletion of SET4 increases sensitivity to acute oxidative stress and alters gene expression patterns (7,(10)(11)(12), indicating a biological function for Set4 even at low abundance. Expression of SET4 appears to be stressregulated, as the transcript and protein levels increase in low oxygen, including hypoxic or anaerobic conditions (13,14).…”

Section: Introductionmentioning

confidence: 99%

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Set4 coordinates the activity of histone deacetylases and regulates stress-responsive gene expression within subtelomeric regions in yeast

Jethmalani

Tran

Jaiswal

et al. 2021

Preprint

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The yeast chromatin protein Set4 is a member of the Set3-subfamily of SET domain proteins which play critical roles in the regulation of gene expression in diverse developmental and environmental contexts, although they appear to lack methyltransferase activity. The molecular functions of Set4 are relatively unexplored, likely due to its low abundance in standard growth conditions. We previously reported that Set4 promotes survival during oxidative stress and regulates expression of stress response genes via stress-dependent chromatin localization. In this study, global gene expression analysis and investigation of histone modification status has revealed a role for Set4 in maintaining gene repressive mechanisms within yeast subtelomeres under both normal and stress conditions. We show that Set4 works in a partially overlapping pathway to the SIR complex and the histone deacetylase Rpd3 to maintain proper levels of histone acetylation and expression of stress response genes encoded in subtelomeres. This role for Set4 is particularly critical for cells under hypoxic conditions, and the loss of Set4 decreases cell fitness and cell wall integrity in hypoxia. These findings uncover a new regulator of subtelomeric chromatin that is key to stress defense pathways and demonstrate a function for yeast Set4 in regulating repressive, heterochromatin-like environments.

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Section: Resultssupporting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Set4 coordinates the activity of histone deacetylases and regulates stress-responsive gene expression within subtelomeric regions in yeast

Jethmalani

Tran

Jaiswal

et al. 2021

Preprint

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“…A drop (5 µl) of each culture was applied on YPD plates and incubated at 30°C (unstressed), 37°C (HTS) and 15°C (low-temperature stress; Mishra et al, 2018). In a separate set of experiments, the cultures were grown on YPD plates independently supplemented with 0.5 M NaCl (hypersalinity; Piao et al, 1999) and 1.2 M sorbitol (osmotic stress; Zhi et al, 2013), and incubated at 30°C for 36 h. For oxidative stress, overnight-grown YPD culture was supplemented with 4 mM H 2 O 2 for 30 min, and similar parameters were followed for spotting on YPD plates (Tran and Green, 2019).…”

Section: Expression Of Ta2cp In Heterologous Systemsmentioning

confidence: 99%

Wheat 2‐Cys peroxiredoxin plays a dual role in chlorophyll biosynthesis and adaptation to high temperature

et al. 2021

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The molecular mechanism of high-temperature stress (HTS) response, in plants, has so far been investigated using transcriptomics, while the dynamics of HTS-responsive proteome remain unexplored. We examined the adaptive responses of the resilient wheat cultivar 'Unnat Halna' and dissected the HTS-responsive proteome landscape. This led to the identification of 55 HTS-responsive proteins (HRPs), which are predominantly involved in metabolism and defense pathways. Interestingly, HRPs included a 2-cysteine peroxiredoxin (2CP), designated Ta2CP, presumably involved in stress perception and adaptation. Complementation of Ta2CP in yeast and heterologous expression in Arabidopsis demonstrated its role in thermotolerance. Both Ta2CP silencing and overexpression inferred the involvement of Ta2CP in plant growth and chlorophyll biosynthesis. We demonstrated that Ta2CP interacts with protochlorophyllide reductase b, TaPORB. Reduced TaPORB expression was found in Ta2cp-silenced plants, while upregulation was observed in Ta2CP-overexpressed plants. Furthermore, the downregulation of Ta2CP in Taporb-silenced plants and reduction of protochlorophyllide in Ta2cp-silenced plants suggested the key role of Ta2CP in chlorophyll metabolism. Additionally, the transcript levels of AGPase1 and starch were increased in Ta2cp-silenced plants. More significantly, HTS-treated Ta2cp-silenced plants showed adaptive responses despite increased reactive oxygen species and peroxide concentrations, which might help in rapid induction of high-temperature acclimation.

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“…The impact of UV-C-induced oxidative stress on DBY746 yeast was examined to test this hypothesis using our published protocol [6,23,26]. The resulting yeast survival, estimated with a quantitative colony counting assay [49], and ROS/RNS production are presented in Figure 4. Under these oxidative conditions induced by UV-C, AE and CHL supplementations significantly improved the survival of yeast (Figure 4a) in conjunction with their ability to Peer-reviewed version available at Life 2020, 10, 80; doi:10.3390/life10060080 significantly reduce ROS / RNS production (Figure 4b).…”

Section: Almond Skin Extract and Chlorogenic Acid Increased Yeast Surmentioning

confidence: 99%

Almond Skin Extracts and Chlorogenic Acid Delay Replicative Aging by Enhanced Oxidative Stress Response Involving SIR2 and SOD2 in Yeast

Abid¹,

Elamrani²,

Drouet³

et al. 2020

Preprint

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Almond (Prunus dulcis (Mill.) D.A.Webb) is one of the largest nut crops in the world. Recently, phenolic compounds, mostly stored in almond skin, have been associated with much of the health-promoting behavior associated with their intake. The almond skin enriched fraction obtained from cold-pressed oil residues of the endemic Moroccan Beldi ecotypes is particularly rich in chlorogenic acid. In this study, both almond skin extract (AE) and chlorogenic acid (CHL) supplements, similar to traditional positive control resveratrol, significantly increased the replicative life-span of yeast compared to the untreated group. Our results showed that AE and CHL significantly reduced the production of reactive oxygen and nitrogen species (ROS/RNS), most likely due to their ability to maintain mitochondrial function during aging, as indicated by the maintenance of normal mitochondrial membrane potential in treated groups. This may be associated with the observed activation of the anti-oxidative stress response in treated yeast, which results in activation at both gene expression and enzymatic activity levels for SOD2 and SIR2, the latter being an upstream inducer of SOD2 expression. Interestingly, the differential gene expression induction of mitochondrial SOD2 gene at the expense of the cytosolic SOD1 gene confirms the key role of mitochondrial function in this regulation. Furthermore, AE and CHL have contributed to the survival of yeast under UV-C-induced oxidative stress, by reducing the development of ROS / RNS, resulting in a significant reduction in cellular oxidative damage as evidenced by decreased membrane lipid peroxidation, protein carbonyl content and 8-oxo-guanine formation in DNA. Together, these results demonstrate the interest of AE and CHL as new regulators in the replicative life-span and control of the oxidative stress response of yeast.

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Assessing Yeast Cell Survival Following Hydrogen Peroxide Exposure

Cited by 25 publications

References 10 publications

Set4 coordinates the activity of histone deacetylases and regulates stress-responsive gene expression within subtelomeric regions in yeast

Set4 coordinates the activity of histone deacetylases and regulates stress-responsive gene expression within subtelomeric regions in yeast

Wheat 2‐Cys peroxiredoxin plays a dual role in chlorophyll biosynthesis and adaptation to high temperature

Almond Skin Extracts and Chlorogenic Acid Delay Replicative Aging by Enhanced Oxidative Stress Response Involving SIR2 and SOD2 in Yeast

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