Adams M R, Sears R, Nuckolls F, Leone G, Nevins J R
Department of Genetics, Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710, USA.
Mol Cell Biol. 2000 May;20(10):3633-9. doi: 10.1128/MCB.20.10.3633-3639.2000.
E2F transcription activity has been shown to play a critical role in cell growth control, regulating the expression of a variety of genes that encode proteins important for the initiation of DNA replication and cell cycle regulation. We have shown that the E2F3 locus encodes two protein products: the E2F3a product, which is tightly regulated by cell growth, and the E2F3b product, which is constitutively expressed throughout the cell cycle. To further explore the mechanism controlling the expression of the two E2F3 gene products, we analyzed the genomic sequences flanking the 5' region of E2F3a and E2F3b. We find that a series of E2F binding sites confer negative control on the E2F3a promoter in quiescent cells, similar to the control of the E2F1 and E2F2 promoters. In addition, a group of E-box elements, which are Myc binding sites, confer responsiveness to Myc and are necessary for full activation of the E2F3a promoter in response to growth stimulation. Based on these results and past experiments, it appears that the E2F1, E2F2, and E2F3a genes are similarly regulated by growth stimulation, involving a combination of E2F-dependent negative control and Myc-mediated positive control. In contrast, the constitutive expression of the E2F3b gene more closely reflects the control of expression of the E2F4 and E2F5 genes.
E2F转录活性已被证明在细胞生长控制中起关键作用,它调控着多种基因的表达,这些基因编码对DNA复制起始和细胞周期调控至关重要的蛋白质。我们已经证明E2F3基因座编码两种蛋白质产物:E2F3a产物,其受细胞生长严格调控;以及E2F3b产物,其在整个细胞周期中持续表达。为了进一步探究控制这两种E2F3基因产物表达的机制,我们分析了E2F3a和E2F3b 5'区域侧翼的基因组序列。我们发现,一系列E2F结合位点在静止细胞中对E2F3a启动子施加负调控,类似于对E2F1和E2F2启动子的调控。此外,一组作为Myc结合位点的E盒元件赋予对Myc的反应性,并且是E2F3a启动子在生长刺激下完全激活所必需的。基于这些结果和以往的实验,似乎E2F1、E2F2和E2F3a基因受到生长刺激的类似调控,涉及E2F依赖性负调控和Myc介导的正调控的组合。相比之下,E2F3b基因的组成性表达更紧密地反映了E2F4和E2F5基因表达的调控。