Single-particle imaging of stress-promoters induction reveals the interplay between MAPK signaling, chromatin and transcription factors

Wosika, Victoria; Pelet, Serge

doi:10.1038/s41467-020-16943-w

Cited by 14 publications

(17 citation statements)

References 67 publications

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“… 2013 ) and it depends on the state of chromatin (Pelet et al . 2011 , Wosika and Pelet 2020 ). Hog1 is a master protein for reprogramming gene expression in response to osmostress through various transcription factors (Rep et al .…”

Section: Osmostress-adaptive Responsesmentioning

confidence: 99%

The HOG pathway and the regulation of osmoadaptive responses in yeast

Nadal

Posas

2022

FEMS Yeast Research

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Cells coordinate intracellular activities in response to changes in the extracellular environment to maximize their probability of survival and proliferation. Eukaryotic cells need to adapt to constant changes in the osmolarity of their environment. In yeast, the HOG (High Osmolarity Glycerol) pathway is responsible for the response to high osmolarity. Activation of the Hog1 stress-activated protein kinase (SAPK) induces a complex program required for cellular adaptation that includes temporary arrest of cell cycle progression, adjustment of transcription and translation patterns, the regulation of metabolism including the synthesis and retention of the compatible osmolyte glycerol. Hog1 is a member of the family of p38 SAPKs, which are present across eukaryotes. Many of the properties of the HOG pathway and downstream regulted proteins are conserved from yeast to mammals. This review addresses the global view of this signaling pathway in yeast, as well as the contribution of Dr. Hohmann's group to the its understanding.

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Section: Osmostress-adaptive Responsesmentioning

confidence: 99%

The HOG pathway and the regulation of osmoadaptive responses in yeast

Nadal

Posas

2022

FEMS Yeast Research

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“…Two media, SD-full and SD-full + 0.4M NaCl + Alexa680-dextran (D34681, Molecular Probes), were output towards the flow chamber via flow generated by pressure controlled reservoirs (LineUp FlowEZ, Fluigent). These media were mixed in calculated ratios to generate specific changes in flow chamber NaCl concentration by a pulse-width modulation scheme with a duty cycle of 500ms using solenoid valves and an Arduino Uno controller 25,36 . Changes in NaCl concentration over time corresponded proportionally to the Alexa680-dextran dye concentration.…”

Section: Flow Chambermentioning

confidence: 99%

“…As these reporter systems become more widely used, it becomes increasingly important to understand how gene architecture controls the intensity and localization of fluorescently labeled mRNA. Obviously, major efforts have been invested in revealing the influence of promoters on the dynamics and level of transcription 24,25 . Sequences controlling the localization and trafficking of the mRNA molecules have been identified 11,26 and export from the nucleus to the cytoplasm has been monitored 27,28 .…”

Section: Introductionmentioning

confidence: 99%

Implication of polymerase recycling for nascent transcript quantification by live cell imaging

Kindongo,

Lieb,

Skaggs

et al. 2023

Preprint

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Take away− Quantification of stress-induced promoter transcription dynamics using a live assays reporter system displays no dependence of signal intensity with gene length.− Mathematical modeling predicts that the formation of gene loops leading to the recycling of polymerases can explain the observed behavior.− More prevalent polymerase recycling on short genes results in a higher transcriptional output.Transcription enables the production of RNA from a DNA template. Due to the highly dynamic nature of transcription, live-cell imaging methods play a crucial role in measuring the kinetics of this process. For instance, transcriptional bursts have been visualized using fluorescent phage-coat proteins that associate tightly with mRNA stem loops formed on nascent transcripts. To convert the signal emanating from a transcription site into meaningful estimates of transcription dynamics, the influence of various parameters on the measured signal must be evaluated. Here, the effect of gene length on the intensity of the transcription site focus was analyzed. Intuitively, a longer gene can support a larger number of transcribing polymerases, thus leading to an increase in the measured signal. However, measurements of transcription induced by hyper-osmotic stress responsive promoters display independence from gene length. A mathematical model of the stress-induced transcription process suggests that the formation of gene loops that favor the recycling of polymerase from the terminator to the promoter can explain the observed behavior. One experimentally validated prediction from this model is that the amount of mRNA produced from a short gene should be higher than for a long one as the density of active polymerase on the short gene will be increased by polymerase recycling. Our data suggest that this recycling contributes significantly to the expression output from a gene and that polymerase recycling is modulated by the promoter identity or the cellular state.

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“…This study-design was repeated in a recent single-cell study with promoter-reporters for stress-responsive genes e.g. STL1 and GPD1 (Wosika and Pelet 2020 ). Applying the same logic as Westfall et al with anchoring Hog1 to the plasma membrane by a fusion to the C-terminal residues of Ras2, they found that the activity of the promoter of STL1 was abolished while the GPD1 promoter was barely affected.…”

Section: Post-translational Modifications and Their Impact On Glycero...mentioning

confidence: 99%

Yeast osmoregulation – glycerol still in pole position

Blomberg

2022

FEMS Yeast Research

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In response to osmotic dehydration cells sense, signal, alter gene expression and metabolically counterbalance osmotic differences. The main compatible solute/osmolyte that accumulates in yeast cells is glycerol, which is produced from the glycolytic intermediate dihydroxyacetone phosphate. This review covers recent advancements in understanding mechanisms involved in sensing, signaling, cell-cycle delays, transcriptional responses as well as post-translational modifications on key proteins in osmoregulation. The protein kinase Hog1 is a key-player in many of these events, however, there is also a growing body of evidence for important Hog1-independent mechanisms playing vital roles. Several missing links in our understanding of osmoregulation will be discussed and future avenues for research proposed. The review highlights that this rather simple experimental system—salt/sorbitol and yeast—has developed into an enormously potent model system unravelling important fundamental aspects in biology. ‘Striving for an integrative view’ (Stefan Hohmann, 2002)

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Single-particle imaging of stress-promoters induction reveals the interplay between MAPK signaling, chromatin and transcription factors

Cited by 14 publications

References 67 publications

The HOG pathway and the regulation of osmoadaptive responses in yeast

The HOG pathway and the regulation of osmoadaptive responses in yeast

Implication of polymerase recycling for nascent transcript quantification by live cell imaging

Yeast osmoregulation – glycerol still in pole position

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