Polarization of yeast cells under different pheromone induction regimens
Data files
Jul 27, 2024 version files 1.22 MB
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1-StepUp_0-0nM.xlsx
46.41 KB
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10-StepDown_50-5nM.xlsx
73.69 KB
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11-StepDown_50-6nM.xlsx
62.36 KB
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12-StepDown_10-0nM.xlsx
89.36 KB
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13-StepDown_10-5nM.xlsx
54.52 KB
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2-StepUp_0-4nM.xlsx
57.91 KB
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3-StepUp_0-5nM.xlsx
55.50 KB
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4-StepUp_0-6nM.xlsx
81.88 KB
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5-StepUp_0-10nM.xlsx
68.95 KB
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6-StepUp_0-50nM.xlsx
89.13 KB
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7-Incremental_50-10-0nM.xlsx
275.32 KB
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8-StepDown_50-0nM.xlsx
53.94 KB
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9-StepDown_50-4nM.xlsx
50.61 KB
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Data_Information.docx
158.53 KB
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README.md
3.38 KB
Abstract
The yeast Saccharomyces cerevisiae undergoes polarized growth in two different contexts: budding and pheromone-induced mating. A key difference between the two processes is that mating polarization occurs in response to an external stimulus, which can be easily manipulated under laboratory conditions. We exploited this property to test the hypothesis that the underlying polarity circuit is bistable. To make this test we used microfluidics to precisely control the temporal profile of mating pheromone and live-cell imaging to monitor polarization in single living cells. Files posted here contain single cell polarity time course data under different pheromone regimens. These data clearly demonstrate that polarization as a function of pheromone concentration depends on the history of the system, a behavior that is a hallmark of bistability. The data also reveal that polarity is rapidly lost following a sudden loss of pheromone.
README: Polarization of yeast cells under different pheromone induction regimens
https://doi.org/10.5061/dryad.4f4qrfjn0
Data sets posted here were used to support the experimental observations presented in "Multiscale modeling of bistability in the yeast polarity circuit" (Hladyshau, Guan et al., 2024). A separate file, "Data_Information.docx", briefly describes the data and provides details for the computational methods used to analyze the data. Full details for the experimental methods can be found in (Vered 2018) "Memory and bistability in the pheromone response pathway." Ph.D. Dissertation, University of North Carolina at Chapel Hill. It should be noted that the data in the posted files are from analyses and some trials made independently from those for figures appearing in the Vered thesis dissertation (Vered 2018).
Description of the data and file structure
Data were collected for polarity establishment in yeast responding to mating pheromone (α-factor). Experiments were performed using microfluidic devices to precisely control the temporal profile of mating pheromone and live-cell imaging to monitor the polarity process in single living cells. To visualize polarity assembly and disassembly, we tagged the polarity scaffold protein Bem1 with a yomNeonGreen fluorescent protein.
Yeast cells polarize under two contexts. One is in response to pheromone during the G1 phase of the cell cycle, and the other is at bud emergence during the G1 to S transition. To distinguish between the two, we exploited a fluorescently tagged myosin II subunit (Myo1-yomRuby), which localizes to the bud neck during all stages of the cell cycle except G1 ((Bhavsar-Jog and Bi 2017); see Fig 2.4 of (Vered 2018)). We restricted our analysis to pheromone induced-polarization by excluding cells with localized Myo1-yomRuby at the time of pheromone exposure. Cells were dropped from the analysis at the time point where the Myo1 spot appeared and for all subsequent time points.
Strain BY4741-198, which expresses both Bem1-yomNeonGreen and Myo1-yomRuby, was used for all polarity experiments. Details of strain construction and all microfluidic procedures are detailed in Lior Vered’s doctoral dissertation (Vered 2018).
Terminal NaN means the cell has exited G1 and is dropped from the analysis.
Any internal NaN means the cell is out of focus in that frame.
The data headings indicating different dates of the experiments are colored for visual clarity.
No specific color coding was used.
The file structure is provided in Table 1 in the "Data_Information.docx" file.
Sharing/Access information
Relevant references:
Bhavsar-Jog, Y. P. and Bi E. (2017). "Mechanics and regulation of cytokinesis in budding yeast." Semin Cell Dev Biol 66: 107-118. PMCID: PMC5474357
Hladyshau S., Guan K., Nivedita N., Errede B., Tsygankov D., and Elston T.C. (2024) "Multiscale modeling of bistability in the yeast polarity circuit." Submitted.
Tsygankov D., Chu P. H., Chen H., Elston T.C., and Hahn K. (2014). "User-friendly tools for quantifying the dynamics of cellular morphology and intracellular protein clusters." Methods Cell Biol. 123: 409-427. PMCID: PMC4504218
Vered, L. (2018). "Memory and bistability in the pheromone response pathway." Ph.D. Dissertation, University of North Carolina at Chapel Hill.
Methods
The accompanying "Data_Information" document explains how polarization was visualized in living cells, the metric used to quantify polarization and a Table summarizing the pheromone regimens under which the data were collected.