A new enrichment approach identifies genes that alter cell cycle progression in Saccharomyces cerevisiae
A new enrichment approach identifies genes that alter cell cycle progression in Saccharomyces cerevisiae
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DOI：
10.1007/s00294-004-0497-5
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发表时间：
2004-06-01
期刊：
影响因子：
2.5
通讯作者：
Polymenis, M
中科院分区：
文献类型：
--
作者：
Bogomolnaya, LM;Pathak, R;Polymenis, M
协调细胞生长与分裂的机制被认为决定了细胞分裂起始的时间并限制了整体细胞增殖。为了确定在酿酒酵母中参与这一过程的基因，我们描述了一种方法，不依赖于细胞大小的改变或对信息素的抗性。相反，我们的方法是基于Flo 1 p蛋白在通过START的细胞中的细胞表面沉积。我们发现，HXT 11（编码质膜转运蛋白），PPE 1（编码蛋白质甲基酯酶）或SIK 1（编码参与rRNA加工的蛋白质）的过度表达缩短了细胞周期G1期的持续时间，在DNA复制开始之前。此外，我们发现，尽管SIK 1不是有丝分裂检查点的一部分，但SIK 1过表达引起纺锤体方向缺陷并致敏G2/M检查点突变细胞。因此，与HXT 11和PPE 1不同，SIK 1过表达也与有丝分裂功能相关。总的来说，我们使用了一种新的富集方法，并确定了以前与细胞周期进展无关的基因。这种方法可以推广到其他生物。
Mechanisms that coordinate cell growth with division are thought to determine the timing of initiation of cell division and to limit overall cell proliferation. To identify genes involved in this process in Saccharomyces cerevisiae, we describe a method that does not rely on cell size alterations or resistance to pheromone. Instead, our approach was based on the cell surface deposition of the Flo1p protein in cells having passed START. We found that over-expression of HXT11 (which encodes a plasma membrane transporter), PPE1 (coding for a protein methyl esterase), or SIK1 (which encodes a protein involved in rRNA processing) shortened the duration of the G1 phase of the cell cycle, prior to the initiation of DNA replication. In addition, we found that, although SIK1 was not part of a mitotic checkpoint, SIK1 over-expression caused spindle orientation defects and sensitized G2/M checkpoint mutant cells. Thus, unlike HXT11 and PPE1, SIK1 over-expression is also associated with mitotic functions. Overall, we used a novel enrichment approach and identified genes that were not previously associated with cell cycle progression. This approach can be extended to other organisms.