A Quantitative Imaging-Based Protocol for Yeast Growth and Survival on Agar Plates

Petropavlovskiy, Andrey A.; Tauro, Michael G.; Lajoie, Patrick; Duennwald, Martin L.

doi:10.1016/j.xpro.2020.100182

Cited by 43 publications

(54 citation statements)

References 10 publications

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“…Percent input from at least three biological replicates is shown. (B) Quantitation of yeast spot assays shown in Fig 7A was performed as described ( Petropavlovskiy et al, 2020 ). The mean gray value taken from the images of plates for at least three biological replicates is shown for each strain and under aerobic or hypoxic conditions.…”

Section: Resultsmentioning

confidence: 99%

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Set4 regulates stress response genes and coordinates histone deacetylases within yeast subtelomeres

et al. 2021

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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. 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 identified 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, where the loss of Set4 decreases cell fitness and cell wall integrity. These findings uncover a new regulator of subtelomeric chromatin that is key to stress defense pathways and demonstrate a function for Set4 in regulating repressive, heterochromatin-like environments.

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Section: Resultsmentioning

confidence: 99%

“…The plates were observed and imaged for 2 d for aerobic conditions and 8 d for hypoxic conditions. The data were quantified as described ( Petropavlovskiy et al, 2020 ). Briefly, the mean gray value quantitation was performed using ImageJ.…”

Section: Methodsmentioning

confidence: 99%

Set4 regulates stress response genes and coordinates histone deacetylases within yeast subtelomeres

et al. 2021

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“…A minimum of three biological replicas were used for quantification. A detailed protocol for the quantification of yeast growth assays is provided in Petropavlovskiy et al, 2020 46 …”

Section: Methodsmentioning

confidence: 99%

Hsp90 and its co‐chaperone Sti1 control TDP‐43 misfolding and toxicity

et al. 2021

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Protein misfolding is a central feature of most neurodegenerative diseases. Molecular chaperones can modulate the toxicity associated with protein misfolding, but it remains elusive which molecular chaperones and co-chaperones interact with specific misfolded proteins. TDP-43 misfolding and inclusion formation are a hallmark of amyotrophic lateral sclerosis (ALS) and other neurodegenerative diseases. Using yeast and mammalian neuronal cells we find that Hsp90 and its co-chaperone Sti1 have the capacity to alter TDP-43 misfolding, inclusion formation, aggregation, and cellular toxicity. Our data also demonstrate that impaired Hsp90 function sensitizes cells to TDP-43 toxicity and that Sti1 specifically interacts with and strongly modulates TDP-43 toxicity in a dose-dependent manner. Our study thus uncovers a previously unrecognized tie between Hsp90, Sti1, TDP-43 misfolding, and cellular toxicity.

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“…Plates were incubated at indicated temperatures for 3 days and photographed using a UVIDOC HD6 camera (Uvitec, Cambridge, United Kingdom). Relative yeast growth on agar plates was quantified by densitometry as described in reference 40 .…”

Section: Methodsmentioning

confidence: 99%

Interplay of the RNA Exosome Complex and RNA-Binding Protein Ssd1 in Maintaining Cell Wall Stability in Yeast

et al. 2021

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A Quantitative Imaging-Based Protocol for Yeast Growth and Survival on Agar Plates

Cited by 43 publications

References 10 publications

Set4 regulates stress response genes and coordinates histone deacetylases within yeast subtelomeres

Set4 regulates stress response genes and coordinates histone deacetylases within yeast subtelomeres

Hsp90 and its co‐chaperone Sti1 control TDP‐43 misfolding and toxicity

Interplay of the RNA Exosome Complex and RNA-Binding Protein Ssd1 in Maintaining Cell Wall Stability in Yeast

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