Role for E2F in control of both DNA replication and mitotic functions as revealed from DNA microarray analysis
Role for E2F in control of both DNA replication and mitotic functions as revealed from DNA microarray analysis
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DOI：
10.1128/mcb.21.14.4684-4699.2001
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发表时间：
2001-07-01
影响因子：
5.3
通讯作者：
Nevins, JR
中科院分区：
文献类型：
--
作者：
Ishida, S;Huang, E;Nevins, JR
我们使用高密度DNA微阵列来分析哺乳动物细胞周期中的基因调控和E2F在这一过程中的作用，细胞周期分析是通过对血清刺激成纤维细胞和羟基脲阻滞在G(1)/S同步的细胞的基因控制的联合检查来促进的，然后释放进入细胞周期。后一种方法（G(1)/S同步）对于严格维持细胞同步对于细胞周期后期基因调控的明确分析至关重要。对这些样本的分析确定了七个不同的基因簇，它们表现出独特的表达模式。基因倾向于根据共同的功能和细胞周期中需要活动的时间聚集在这些组中。在这种情况下，对E2F蛋白诱导的基因的分析鉴定了以前未被E2F蛋白调节的基因或表达序列标签；令人惊讶的是，其中许多编码的蛋白质在有丝分裂过程中起作用。对e2f诱导基因与细胞生长调节基因表达模式的比较显示，几乎所有的e2f诱导基因仅在两个细胞周期簇中发现；其中一组在G(1)/S处受到调控，第二组包括有丝分裂活性在内在G(2)处受到调控。G(2)基因的激活表明E2F在控制DNA复制和有丝分裂活性方面具有更广泛的作用。
We have used high-density DNA microarrays to provide an analysis of gene regulation during the mammalian cell cycle and the role of E2F in this process, Cell cycle analysis was facilitated by a combined examination of gene control in serum-stimulated fibroblasts and cells synchronized at G(1)/S by hydroxyurea block that were then released to proceed through the cell cycle. The latter approach (G(1)/S synchronization) is critical for rigorously maintaining cell synchrony for unambiguous analysis of gene regulation in later stages of the cell cycle. Analysis of these samples identified seven distinct clusters of genes that exhibit unique patterns of expression. Genes tend to cluster within these groups based on common function and the time during the cell cycle that the activity is required. Placed in this context, the analysis of genes induced by E2F proteins identified genes or expressed sequence tags not previously described as regulated by E2F proteins; surprisingly, many of these encode proteins known to function during mitosis. A comparison of the E2F-induced genes with the patterns of cell growth-regulated gene expression revealed that virtually all of the E2F-induced genes are found in only two of the cell cycle clusters; one group was regulated at G(1)/S, and the second group, which included the mitotic activities, was regulated at G(2). The activation of the G(2) genes suggests a broader role for E2F in the control of both DNA replication and mitotic activities.