Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
Temporal Expression of a Master Regulator Drives Synchronous Sporulation in Budding Yeast
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DOI：
10.1534/g3.116.034983
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发表时间：
2016-11-01
影响因子：
2.6
通讯作者：
van Werven, Folkert J.
中科院分区：
文献类型：
--
作者：
Chia, Minghao;van Werven, Folkert J.
酵母细胞进入和经历配子体发生是相对不同步的，这使得进行特定阶段的基因组和生化分析在技术上具有挑战性。进入孢子形成的主要调控因子IME1在细胞间的表达差异被认为是不同步孢子形成的潜在原因。在这里，我们发现IME1表达的时机对于诱导细胞同步进行孢子形成至关重要。当我们在孢子培养基中培养2小时的细胞中诱导启动子表达IME1时，绝大多数细胞在减数分裂前DNA复制和减数分裂过程中表现出同步。诱导IME1表达过早或过晚都会影响产孢的同步性。令人惊讶的是，我们的同步产孢方法不需要在含醋酸盐的培养基中生长，但可以在富培养基中生长直至饱和的细胞中实现。我们的系统只需要IME1，因为n6 -甲基腺苷甲基转移酶IME4的表达是由IME1本身控制的，而IME4是早期孢子形成的另一个关键调节因子。这里描述的方法可以很容易地与其他特定阶段的同步方法相结合，从而应用于研究孢子形成的特定阶段，或完整的孢子形成程序。
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.