Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast

Yeast cells enter and undergo gametogenesis relatively asynchronously, making it technically challenging to perform stage-specific genomic and biochemical analyses. Cell-to-cell variation in the expression of the master regulator of entry into sporulation, IME1, has been implicated to be the underly...

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Main Authors: Minghao Chia, Folkert J. van Werven
Format: Article
Language:English
Published: Oxford University Press 2016-11-01
Series:G3: Genes, Genomes, Genetics
Subjects:
Online Access:http://g3journal.org/lookup/doi/10.1534/g3.116.034983
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author Minghao Chia
Folkert J. van Werven
author_facet Minghao Chia
Folkert J. van Werven
author_sort Minghao Chia
collection DOAJ
description Yeast cells enter and undergo gametogenesis relatively asynchronously, making it technically challenging to perform stage-specific genomic and biochemical analyses. Cell-to-cell variation in the expression of the master regulator of entry into sporulation, IME1, has been implicated to be the underlying cause of asynchronous sporulation. Here, we find that timing of IME1 expression is of critical importance for inducing cells to undergo sporulation synchronously. When we force expression of IME1 from an inducible promoter in cells incubated in sporulation medium for 2 hr, the vast majority of cells exhibit synchrony during premeiotic DNA replication and meiotic divisions. Inducing IME1 expression too early or too late affects the synchrony of sporulation. Surprisingly, our approach for synchronous sporulation does not require growth in acetate-containing medium, but can be achieved in cells grown in rich medium until saturation. Our system requires solely IME1, because the expression of the N6-methyladenosine methyltransferase IME4, another key regulator of early sporulation, is controlled by IME1 itself. The approach described here can be combined easily with other stage-specific synchronization methods, and thereby applied to study specific stages of sporulation, or the complete sporulation program.
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spelling doaj.art-1b629aae24c3450aadba30f6af45d9d12022-12-21T22:22:36ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362016-11-016113553356010.1534/g3.116.03498312Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding YeastMinghao ChiaFolkert J. van WervenYeast cells enter and undergo gametogenesis relatively asynchronously, making it technically challenging to perform stage-specific genomic and biochemical analyses. Cell-to-cell variation in the expression of the master regulator of entry into sporulation, IME1, has been implicated to be the underlying cause of asynchronous sporulation. Here, we find that timing of IME1 expression is of critical importance for inducing cells to undergo sporulation synchronously. When we force expression of IME1 from an inducible promoter in cells incubated in sporulation medium for 2 hr, the vast majority of cells exhibit synchrony during premeiotic DNA replication and meiotic divisions. Inducing IME1 expression too early or too late affects the synchrony of sporulation. Surprisingly, our approach for synchronous sporulation does not require growth in acetate-containing medium, but can be achieved in cells grown in rich medium until saturation. Our system requires solely IME1, because the expression of the N6-methyladenosine methyltransferase IME4, another key regulator of early sporulation, is controlled by IME1 itself. The approach described here can be combined easily with other stage-specific synchronization methods, and thereby applied to study specific stages of sporulation, or the complete sporulation program.http://g3journal.org/lookup/doi/10.1534/g3.116.034983gametogenesissporulationsynchronybudding yeastDNA replicationmeiotic divisionsIME1IME4temporal
spellingShingle Minghao Chia
Folkert J. van Werven
Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
G3: Genes, Genomes, Genetics
gametogenesis
sporulation
synchrony
budding yeast
DNA replication
meiotic divisions
IME1
IME4
temporal
title Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
title_full Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
title_fullStr Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
title_full_unstemmed Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
title_short Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
title_sort temporal expression of a master regulator drives synchronous sporulation in budding yeast
topic gametogenesis
sporulation
synchrony
budding yeast
DNA replication
meiotic divisions
IME1
IME4
temporal
url http://g3journal.org/lookup/doi/10.1534/g3.116.034983
work_keys_str_mv AT minghaochia temporalexpressionofamasterregulatordrivessynchronoussporulationinbuddingyeast
AT folkertjvanwerven temporalexpressionofamasterregulatordrivessynchronoussporulationinbuddingyeast