Raithatha Sheetal A, Stuart David T
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Genetics. 2005 Mar;169(3):1329-42. doi: 10.1534/genetics.104.036103. Epub 2005 Jan 16.
In proliferating S. cerevisiae, genes whose products function in DNA replication are regulated by the MBF transcription factor composed of Mbp1 and Swi6 that binds to consensus MCB sequences in target promoters. We find that during meiotic development a subset of DNA replication genes exemplified by TMP1 and RNR1 are regulated by Mbp1. Deletion of Mbp1 deregulated TMP1 and RNR1 but did not interfere with premeiotic S-phase, meiotic recombination, or spore formation. Surprisingly, deletion of MBP1 had no effect on the expression of CLB5, which is purportedly controlled by MBF. Extensive analysis of the CLB5 promoter revealed that the gene is largely regulated by elements within a 100-bp fragment containing a cluster of MCB sequences. Surprisingly, induction of the CLB5 promoter requires MCB sequences, but not Mbp1, implying that another MCB-binding factor may exist in cells undergoing meiosis. In addition, full activation of CLB5 during meiosis requires Clb5 activity, suggesting that CLB5 may be regulated by a positive feedback mechanism. We further demonstrate that during meiosis MCBs function as effective transcriptional activators independent of MBP1.
在增殖的酿酒酵母中,其产物在DNA复制中起作用的基因受由Mbp1和Swi6组成的MBF转录因子调控,该转录因子与靶启动子中的共有MCB序列结合。我们发现,在减数分裂发育过程中,以TMP1和RNR1为代表的一部分DNA复制基因受Mbp1调控。Mbp1的缺失使TMP1和RNR1的表达失调,但不干扰减数分裂前的S期、减数分裂重组或孢子形成。令人惊讶的是,MBP1的缺失对CLB5的表达没有影响,而CLB5据称是由MBF控制的。对CLB5启动子的广泛分析表明,该基因在很大程度上受一个包含MCB序列簇的100 bp片段内元件的调控。令人惊讶的是,CLB5启动子的诱导需要MCB序列,但不需要Mbp1,这意味着在减数分裂细胞中可能存在另一种MCB结合因子。此外,减数分裂期间CLB5的完全激活需要Clb5活性,这表明CLB5可能受正反馈机制调控。我们进一步证明,在减数分裂过程中,MCB作为独立于MBP1的有效转录激活因子发挥作用。