Maher M, Cong F, Kindelberger D, Nasmyth K, Dalton S
Roche Institute of Molecular Biology, Nutley, New Jersey 07110-1199, USA.
Mol Cell Biol. 1995 Jun;15(6):3129-37. doi: 10.1128/MCB.15.6.3129.
Clb2 is the major B-type mitotic cyclin required for entry into mitosis in the budding yeast Saccharomyces cerevisiae. We showed that accumulation of CLB2 transcripts in G2 cells is controlled at the transcriptional level and identified a 55-bp upstream activating sequence (UAS) containing an Mcm1 binding site as being necessary and sufficient for cell cycle regulation. Sequences within the cell cycle-regulated UAS were shown to bind Mcm1 in vitro, and mutation which abolished Mcm1-dependent DNA binding activity eliminated cell cycle-regulated transcription in vivo. A second protein with no autonomous DNA binding activity was also recruited into Mcm1-UAS complexes, generating a ternary complex. A point mutation in the CLB2 UAS which blocked ternary complex formation, but still allowed Mcm1 to bind, resulted in loss of cell cycle regulation in vivo, suggesting that the ternary complex factor is also important in control of CLB2 transcription. We discuss the possibility that the CLB2 gene is coregulated with other genes known to be regulated with the same periodicity and suggest that Mcm1 and the ternary complex factor may coordinately regulate several other G2-regulated transcripts.
Clb2是芽殖酵母酿酒酵母进入有丝分裂所需的主要B型有丝分裂周期蛋白。我们发现G2期细胞中CLB2转录本的积累在转录水平受到调控,并鉴定出一个含有Mcm1结合位点的55碱基对上游激活序列(UAS),它对于细胞周期调控是必要且充分的。细胞周期调控的UAS内的序列在体外显示能结合Mcm1,而消除Mcm1依赖性DNA结合活性的突变在体内消除了细胞周期调控的转录。一种没有自主DNA结合活性的第二种蛋白质也被招募到Mcm1-UAS复合物中,形成三元复合物。CLB2 UAS中的一个点突变阻止了三元复合物的形成,但仍允许Mcm1结合,导致体内细胞周期调控丧失,这表明三元复合物因子在CLB2转录控制中也很重要。我们讨论了CLB2基因与已知以相同周期调控的其他基因共同调控的可能性,并提出Mcm1和三元复合物因子可能协同调控其他几个G2期调控的转录本。