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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Genetics Society of America
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5100854/ https://www.ncbi.nlm.nih.gov/pubmed/27605516 http://dx.doi.org/10.1534/g3.116.034983 |
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author | Chia, Minghao van Werven, Folkert J. |
author_facet | Chia, Minghao van Werven, Folkert J. |
author_sort | Chia, Minghao |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-5100854 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Genetics Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-51008542016-11-09 Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast Chia, Minghao van Werven, Folkert J. G3 (Bethesda) Investigations 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. Genetics Society of America 2016-09-06 /pmc/articles/PMC5100854/ /pubmed/27605516 http://dx.doi.org/10.1534/g3.116.034983 Text en Copyright © 2016 Chia, and van Werven http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Investigations Chia, Minghao van Werven, Folkert J. Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast |
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 | Investigations |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5100854/ https://www.ncbi.nlm.nih.gov/pubmed/27605516 http://dx.doi.org/10.1534/g3.116.034983 |
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