Bogomolnaya Lydia M, Pathak Ritu, Cham Roxana, Guo Jinbai, Surovtseva Yulia V, Jaeckel Lane, Polymenis Michael
Department of Biochemistry and Biophysics, Texas A&M University, 2128 TAMU, College Station, TX 77843, USA.
Curr Genet. 2004 Jun;45(6):350-9. doi: 10.1007/s00294-004-0497-5. Epub 2004 Mar 12.
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.
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,并限制整体细胞增殖。为了鉴定酿酒酵母中参与这一过程的基因,我们描述了一种不依赖细胞大小改变或对信息素抗性的方法。相反,我们的方法基于Flo1p蛋白在通过START的细胞中的细胞表面沉积。我们发现,HXT11(编码一种质膜转运蛋白)、PPE1(编码一种蛋白质甲基酯酶)或SIK1(编码一种参与rRNA加工的蛋白质)的过表达缩短了DNA复制起始前细胞周期G1期的持续时间。此外,我们发现,虽然SIK1不是有丝分裂检查点的一部分,但SIK1的过表达会导致纺锤体定向缺陷,并使G2/M检查点突变细胞敏感。因此,与HXT11和PPE1不同,SIK1的过表达也与有丝分裂功能有关。总体而言,我们使用了一种新的富集方法,并鉴定了以前与细胞周期进程无关的基因。这种方法可以扩展到其他生物体。