Mechanisms of cell cycle control revealed by a systematic and quantitative overexpression screen in S. cerevisiae.
Mechanisms of cell cycle control revealed by a systematic and quantitative overexpression screen in S. cerevisiae.
复制标题
DOI：
10.1371/journal.pgen.1000120
复制
发表时间：
2008-07-11
期刊：
影响因子：
4.5
通讯作者：
Marcotte EM
中科院分区：
文献类型：
--
作者：
Niu W;Li Z;Zhan W;Iyer VR;Marcotte EM
细胞周期进程的调节是细胞健康和繁殖的基础，这一过程的失败与许多人类疾病有关。我们对细胞周期调节因子的了解大多来自于功能丧失的研究。为了揭示在功能丧失研究中难以识别的新的细胞周期调控基因，我们对酵母基因过表达诱导的细胞周期延迟表型进行了近基因组范围的流式细胞术测定。我们确定了108个基因，其过表达显著延迟了酵母细胞周期在特定阶段的进展。许多新的基因与细胞周期进程有关，例如SKO 1、RFA 1和YPR 015 C。RFA 1或YPR 015 C的过表达分别通过破坏纺锤体与染色体的附着和激活DNA损伤检查点而使细胞周期延迟在G2/M期。与此相反，过度表达的转录因子SKO 1逮捕细胞在G1期通过激活信息素反应途径，揭示了新的串扰之间的渗透感应和交配。更一般地，92%-94%的基因在过表达时与其相应的缺失突变体相比表现出不同的表型，支持许多基因可以在过表达时获得功能的观点。因此，这项工作暗示了细胞周期进程中的新基因，补充了之前的筛选，并为未来的实验奠定了基础，以更精确地定义这些基因在细胞周期进程中的作用。所有细胞都需要适当的细胞周期调节;失败会导致许多人类疾病。细胞周期机制在真核生物中广泛保守，已知许多关键调控基因。尽管如此，我们对监管机构的了解是不完整的。许多经典的研究分析酵母功能丧失突变体，以确定细胞周期基因。研究还涉及基于其过表达表型的基因，但基因过表达对细胞周期的影响尚未被量化的所有酵母基因。我们单独量化了几乎所有（91%）酵母基因的过表达对细胞周期进程的影响，我们报告了108个导致最显著和可重复的细胞周期缺陷的基因，其中大部分以前没有观察到。我们更详细地描述了三个基因，其中一个涉及染色体分离和有丝分裂纺锤体的形成。第二个影响有丝分裂稳定性和DNA损伤检查点。奇怪的是，第三个基因SKO 1的过度表达通过激活信息素反应途径来阻止细胞周期，细胞错误地表现为交配信息素存在。这些结果为进一步阐明这些基因在细胞周期中的作用奠定了基础。在人类细胞中进行类似的试验可以帮助进一步阐明细胞周期控制和癌症之间的许多联系。
Regulation of cell cycle progression is fundamental to cell health and reproduction, and failures in this process are associated with many human diseases. Much of our knowledge of cell cycle regulators derives from loss-of-function studies. To reveal new cell cycle regulatory genes that are difficult to identify in loss-of-function studies, we performed a near-genome-wide flow cytometry assay of yeast gene overexpression-induced cell cycle delay phenotypes. We identified 108 genes whose overexpression significantly delayed the progression of the yeast cell cycle at a specific stage. Many of the genes are newly implicated in cell cycle progression, for example SKO1, RFA1, and YPR015C. The overexpression of RFA1 or YPR015C delayed the cell cycle at G2/M phases by disrupting spindle attachment to chromosomes and activating the DNA damage checkpoint, respectively. In contrast, overexpression of the transcription factor SKO1 arrests cells at G1 phase by activating the pheromone response pathway, revealing new cross-talk between osmotic sensing and mating. More generally, 92%–94% of the genes exhibit distinct phenotypes when overexpressed as compared to their corresponding deletion mutants, supporting the notion that many genes may gain functions upon overexpression. This work thus implicates new genes in cell cycle progression, complements previous screens, and lays the foundation for future experiments to define more precisely roles for these genes in cell cycle progression. All cells require proper cell cycle regulation; failure leads to numerous human diseases. Cell cycle mechanisms are broadly conserved across eukaryotes, with many key regulatory genes known. Nonetheless, our knowledge of regulators is incomplete. Many classic studies have analyzed yeast loss-of-function mutants to identify cell cycle genes. Studies have also implicated genes based upon their overexpression phenotypes, but the effects of gene overexpression on the cell cycle have not been quantified for all yeast genes. We individually quantified the effect of overexpression on cell cycle progression for nearly all (91%) of yeast genes, and we report the 108 genes causing the most significant and reproducible cell cycle defects, most of which have not been previously observed. We characterize three genes in more detail, implicating one in chromosomal segregation and mitotic spindle formation. A second affects mitotic stability and the DNA damage checkpoint. Curiously, overexpression of a third gene, SKO1, arrests the cell cycle by activating the pheromone response pathway, with cells mistakenly behaving as if mating pheromone is present. These results establish a basis for future experiments elucidating precise cell cycle roles for these genes. Similar assays in human cells could help further clarify the many connections between cell cycle control and cancers.
登录
查看更多内容
影响因子：
21.3
作者：
Kittler, Ralf;Pelletier, Laurence;Buchholz, Frank
通讯作者：
Buchholz, Frank
影响因子：
16
作者：
Cho, RJ;Campbell, MJ;Davis, RW
通讯作者：
Davis, RW
影响因子：
3.2
作者：
HARTWELL, LH
通讯作者：
HARTWELL, LH
影响因子：
5.8
作者：
de Lichtenberg, U;Jensen, LJ;Brunak, S
通讯作者：
Brunak, S
DOI：
10.1073/pnas.0605140103
发表时间：
2006-08-08
影响因子：
11.1
作者：
Chua, Gordon;Morris, Quaid D.;Hughes, Timothy R.
通讯作者：
Hughes, Timothy R.