Chemical genetic induction of meiosis in Schizosaccharomyces pombe
Chemical genetic induction of meiosis in Schizosaccharomyces pombe
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DOI：
10.4161/cc.20051
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发表时间：
2012-04-15
期刊：
影响因子：
4.3
通讯作者：
Ayte, Jose
中科院分区：
文献类型：
--
作者：
Guerra-Moreno, Angel;Alves-Rodrigues, Isabel;Ayte, Jose
在裂殖酵母裂殖酵母中，减数分裂被蛋白激酶Pat 1抑制，Pat 1磷酸化并灭活Mei 2，Mei 2是减数分裂起始所必需的RNA结合蛋白。当二倍体细胞缺乏营养时，它们启动一系列事件，导致Pat 1失活并进入减数分裂。携带温度敏感的pat 1 -114等位基因的菌株在转换到非允许温度时被迫进入减数分裂，与细胞的倍性无关。该系统已被广泛使用，因为它可以实现高度同步的减数分裂，这是任何旨在破译减数分裂机制的分子或微观方法所必需的。在这里，我们设计了一个新的系统，以获得类似的同步减数分裂，但独立的温度变化。因此，通过在Pat 1的ATP口袋中引入突变，我们已经产生了一种蛋白激酶，该蛋白激酶在小的特异性抑制剂的存在下可以被灭活。这导致在不需要温度变化的情况下强制进入减数分裂，最小化热休克或任何其他应激反应的引入，沿着转录的减数分裂波。
In the fission yeast Schizosaccharomyces pombe, meiosis is inhibited by the protein kinase Pat1, which phosphorylates and inactivates Mei2, an RNA binding protein essential for the initiation of meiosis. When diploid cells are deprived of nutrients, they initiate a cascade of events leading to the inactivation of Pat1 and entry into meiosis. Strains carrying the temperature-sensitive pat1-114 allele are forced to enter into meiosis when shifted to the non-permissive temperature, independently of the ploidity of the cell. This system has been extensively used, since it is possible to achieve a highly synchronous meiosis, which is a must for any molecular or microscopic approach that aims to decipher the mechanisms governing meiosis. Here, we have designed a new system to obtain a similarly synchronous meiosis, but independently of temperature shifts. Thus, by introducing a mutation in the ATP pocket of Pat1, we have generated a protein kinase that, in the presence of small specific inhibitors, can be inactivated. This results in forced entry into meiosis without the need of a temperature shift, minimizing the introduction of heat shock or any other stress responses along the meiotic waves of transcription.