Wade C, Shea K A, Jensen R V, McAlear M A
Molecular Biology and Biochemistry Department, Wesleyan University, Middletown, CT 06459, USA.
Mol Cell Biol. 2001 Dec;21(24):8638-50. doi: 10.1128/MCB.21.24.8638-8650.2001.
In an effort to identify sets of yeast genes that are coregulated across various cellular transitions, gene expression data sets derived from yeast cells progressing through the cell cycle, sporulation, and diauxic shift were analyzed. A partitioning algorithm was used to divide each data set into 24 clusters of similar expression profiles, and the membership of the clusters was compared across the three experiments. A single cluster of 189 genes from the cell cycle experiment was found to share 65 genes with a cluster of 159 genes from the sporulation data set. Many of these genes were found to be clustered in the diauxic-shift experiment as well. The overlapping set was enriched for genes required for rRNA biosynthesis and included genes encoding RNA helicases, subunits of RNA polymerases I and III, and rRNA processing factors. A subset of the 65 genes was tested for expression by a quantitative-relative reverse transcriptase PCR technique, and they were found to be coregulated after release from alpha factor arrest, heat shock, and tunicamycin treatment. Promoter scanning analysis revealed that the 65 genes within this ribosome and rRNA biosynthesis (RRB) regulon were enriched for two motifs: the 13-base GCGATGAGATGAG and the 11-base TGAAAAATTTT consensus sequences. Both motifs were found to be important for promoting gene expression after release from alpha factor arrest in a test rRNA processing gene (EBP2), which suggests that these consensus sequences may function broadly in the regulation of a set of genes required for ribosome and rRNA biosynthesis.
为了确定在各种细胞转变过程中共同调控的酵母基因集，对源自经历细胞周期、孢子形成和双相转变的酵母细胞的基因表达数据集进行了分析。使用一种划分算法将每个数据集划分为24个具有相似表达谱的簇，并在三个实验中比较这些簇的成员。发现细胞周期实验中的一个由189个基因组成的簇与孢子形成数据集中一个由159个基因组成的簇共享65个基因。其中许多基因在双相转变实验中也聚集在一起。重叠集中富含rRNA生物合成所需的基因，包括编码RNA解旋酶、RNA聚合酶I和III的亚基以及rRNA加工因子的基因。通过定量相对逆转录酶PCR技术对65个基因的一个子集进行了表达测试，发现它们在从α因子阻滞、热休克和衣霉素处理中释放后共同调控。启动子扫描分析表明，这个核糖体和rRNA生物合成（RRB）调控子中的65个基因富含两个基序：13碱基的GCGATGAGATGAG和11碱基的TGAAAAATTTT共有序列。在一个测试的rRNA加工基因（EBP2）中，发现这两个基序对于从α因子阻滞中释放后促进基因表达都很重要，这表明这些共有序列可能在核糖体和rRNA生物合成所需的一组基因的调控中广泛发挥作用。
