Althoefer H, Schleiffer A, Wassmann K, Nordheim A, Ammerer G
Institute for Molecular Biology, Hannover Medical School, Germany.
Mol Cell Biol. 1995 Nov;15(11):5917-28. doi: 10.1128/MCB.15.11.5917.
In the budding yeast Saccharomyces cerevisiae, MCM1 encodes an essential DNA-binding protein that regulates transcription of many genes in cooperation with different associated factors. With the help of a conditional expression system, we show that Mcm1 depletion has a distinct effect on cell cycle progression by preventing cells from undergoing mitosis. Genes that normally exhibit a G2-to-M-phase-specific expression pattern, such as CLB1, CLB2, CDC5, SWI5, and ACE2, remain uninduced in the absence of functional Mcm1. In vivo footprinting experiments show that Mcm1, in conjunction with an Mcm1-recruited factor, binds to the promoter regions of SWI5 and CLB2 at sites shown to be involved in cell cycle regulation. However, promoter occupation at these sites is cell cycle independent, and therefore the regulatory system seems to operate on constitutively bound Mcm1 complexes. A gene fusion that provides Mcm1 with a strong transcriptional activation domain causes transcription of SWI5, CLB1, CLB2, and CDC5 at inappropriate times of the cell cycle. Thus, Mcm1 and a cooperating, cell cycle-regulated activation partner are directly involved in the coordinated expression of multiple G2-regulated genes. The arrest phenotype of Mcm1-depleted cells is consistent with low levels of Clb1 and Clb2 kinase. However, constitutive CLB2 expression does not suppress the mitotic defect, and therefore other essential activities required for the G2-to-M transition must also depend on Mcm1 function.
在出芽酵母酿酒酵母中,MCM1编码一种必需的DNA结合蛋白,该蛋白与不同的相关因子协同调节许多基因的转录。借助条件表达系统,我们发现Mcm1缺失通过阻止细胞进行有丝分裂而对细胞周期进程产生独特影响。正常情况下呈现G2期到M期特异性表达模式的基因,如CLB1、CLB2、CDC5、SWI5和ACE2,在缺乏功能性Mcm1时仍未被诱导。体内足迹实验表明,Mcm1与一种被Mcm1招募的因子结合,在显示参与细胞周期调控的位点与SWI5和CLB2的启动子区域结合。然而,这些位点的启动子占据与细胞周期无关,因此调控系统似乎作用于组成型结合的Mcm1复合物。一种为Mcm1提供强转录激活结构域的基因融合导致SWI5、CLB1、CLB2和CDC5在细胞周期的不适当时间转录。因此,Mcm1和一个协同的、受细胞周期调控的激活伙伴直接参与多个G2期调控基因的协调表达。Mcm1缺失细胞的停滞表型与低水平的Clb1和Clb2激酶一致。然而,组成型CLB2表达并不能抑制有丝分裂缺陷,因此G2期到M期转换所需的其他必需活性也必须依赖于Mcm1功能。